CN109746063A - Microlayer model detection system - Google Patents
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- CN109746063A CN109746063A CN201711074989.0A CN201711074989A CN109746063A CN 109746063 A CN109746063 A CN 109746063A CN 201711074989 A CN201711074989 A CN 201711074989A CN 109746063 A CN109746063 A CN 109746063A
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention provides a kind of microlayer model detection system, and the microlayer model detection system includes the first component, second component and third member；The device when first component is microlayer model detection for waste collection after the sample-adding and detection of oil removal and upper oil slick；The second component is microlayer model detection system, the microlayer model detection system includes a centre bore, the centre bore for injected in the microlayer model detection system preparation process during injection molding material and mass production and/or in mass fluorescence detection microlayer model detection system transhipment；The container that microlayer model carries out microlayer model PCR amplification is equipped with the third member.The microlayer model detection system, can it is quick, reliable, convenient and it is free of contamination detection microlayer model sample fluorescence signal；The material and processing cost of the microlayer model detection system are low, are conducive to the extensive use of clinical detection and other biological detection.
Description
Technical field
The present invention relates to microlayer model digital pcr technical fields, and in particular to a kind of microlayer model detection system.
Background technique
Microlayer model digital pcr technology (droplet digital PCR, ddPCR) is a kind of nucleic acid based on single-molecule PCR
Absolute quantification analysis technology.Just to become industry next for highly sensitive, high accuracy advantage with it for microlayer model digital pcr technology
Revolutionary technology.In recent years, with micro-nano manufacturing technology and micro-fluidic technologies (micro-nanofabrication and
Microfluidics development), microlayer model digital pcr technology encounter the best opportunity of break-through skill bottleneck.The technology by
Micro-fluidic chip, generating diameter is a few micrometers of drops for arriving hundreds of microns；Microlayer model wraps up unimolecule or unicellular, reaches reaction
It is totally-enclosed with detecting, it is fully integrated.Microlayer model digital pcr System Working Principle is: generating instrument by special microlayer model first will
Sample to be tested is assigned in " Water-In-Oil " microlayer model of a large amount of nanoliter levels (diameter is a few micrometers to hundreds of microns), the number of microlayer model
Amount is in million ranks.It is mutually isolated by oil reservoir between microlayer model since microlayer model quantity is enough, therefore each microlayer model is suitable
The DNA unimolecule of sample to be tested is contained only in one " microreactor ", microlayer model；Then, turned for these microlayer models
It moves on in EP pipe, is reacted in a Standard PCR instrument.By the microlayer model of pcr amplification reaction, pass through special microlayer model
Analyzer one by one detects the fluorescence signal of drop, and the droplet interpretation for having fluorescence signal is 1, not the droplet of fluorescence signal
Interpretation is 0.Finally, according to the target dna of the number and ratio of Poisson distribution principle and positive droplet you can get it sample to be tested
Molecule amount realizes the absolute quantitation to sample of nucleic acid.One committed step of microlayer model digital pcr technology is quick, reliable
The fluorescence signal of judgement microlayer model sample identify, count the number and ratio of positive microlayer model.
The judgement of microlayer model sample fluorescence signal relies on a core technology: the design of microlayer model fluorescence detection device and adding
Work distinguishes negative microlayer model and positive microlayer model using the fluorescence signal height of product in laser excitation microlayer model.Microlayer model
The old process of digital pcr technology is: the microlayer model of generation is transferred in EP pipe, is reacted in a Standard PCR instrument.
By the microlayer model of pcr amplification reaction, it is injected into a microlayer model fluorescence detection device, special microlayer model is cooperated to analyze
Instrument carries out fluorescence signal detection.The microlayer model fluorescence detection device, which is widely used, needs to have following principle with clinical detection:
(1) the EP pipe containing the microlayer model by pcr amplification reaction is not necessarily to transfer with the microlayer model fluorescence detection device airtight connection,
Reduce possible cross contamination；(2) microlayer model is injected into the microlayer model fluorescence detection under the effect of special microlayer model analyzer
In device；When microlayer model flows through laser detection area, single proper alignment, convenient for the accurate detection of fluorescence signal；(3) after detecting
Microlayer model is collected into a closed storage container, reduces possible cross contamination；(4) the microlayer model fluorescence detection device
Disposable, material and processing cost are low, and (5) are convenient for the operation of the microlayer model fluorescence detection device.For the above original
Then, the microlayer model detection system based on microflow control technique has great application prospect.
Currently, the micro-fluidic chip based on dimethyl silicone polymer (PDMS) has been widely used for detection microlayer model.Firstly,
Researcher has the PDMS microlayer model chip of micron dimension using soft light carving technology (manual operation) processing.When PDMS microlayer model
After chip is successfully prepared, outlet being generated in its sample inlet, microlayer model and is punched using mechanical processing technique, assembly sample feeding pipe goes out
Sample pipe." oily phase " sample, " microlayer model " sample in EP pipe are drawn into syringe by manual mode.Then, pass through outside
Syringe pump is by " oily phase " sample, " microlayer model " samples by sample feeding pipe injection PDMS microlayer model chip.It is being pre-designed
Good flow passage area, Systems for optical inspection one by one detect the fluorescence signal of drop.Finally, detected microlayer model passes through
Sample outlet pipe is collected into routine experiment consumptive material, such as EP pipe.Although PDMS microlayer model chip material researches and develops at low cost, laboratory
Processing technology is simple, but its existing deficiency includes:
(1) PDMS microlayer model detection system manages open connection with the EP containing " microlayer model " sample, be easy to cause intersection
Pollution.
(2) microlayer model after detecting is collected into an open EP pipe, be easy to cause cross contamination.
(3) PDMS is thermoelastic polymer material, such material is not suitable for technical grade injection molding, packaging technology.Add by hand
The PDMS microlayer model chip reliability of work is poor.PDMS microlayer model chip batch processing cost is high.
(4) injection of PDMS microlayer model chip sample, drop are collected as the cumbersome manual operation process of process, are unsuitable for clinic
Examine application.
For the deficiency of PDMS microlayer model chip, we design and machined the microlayer model detection system based on microflow control technique
System.The microlayer model detection system, can it is quick, reliable, convenient and it is free of contamination detection microlayer model sample fluorescence signal；
The material and processing cost of the microlayer model detection system are low, are conducive to the extensive use of clinical detection.
Summary of the invention
In order to overcome the above problem, it includes that the present invention, which provides microlayer model detection system described in a kind of microlayer model detection system,
One component, second component and third member；The sample-adding when first component is microlayer model detection for oil removal and upper oil slick
And detection after waste collection device；The second component is microlayer model detection chip, and the microlayer model detection system includes
There is a centre bore, the centre bore is for injecting injection molding material and mass production in the microlayer model detection system preparation process
The transhipment of microlayer model detection system in the process and/or in mass fluorescence detection；Micro- liquid is equipped with the third member
Drop carries out the container of microlayer model PCR amplification；The second component is fixedly connected on the upper surface of the first component；With described
Three components are securely removably connected at the lower surface of the first component.
In one embodiment, the first component and the second component pass through dispensing mode or ultrasonic welding mode
It is sealedly and fixedly connected.
In one embodiment, the first component and the second component are formed by integrated injection molding respectively.
In one embodiment, the first component and the second component are thermoplastic materials, preferably poly- carbonic acid
Ester material, cyclic olefine copolymer or polymethyl methacrylate, polypropylene.
In one embodiment, the first component is provided with the detent slot for fixing the third member.
In one embodiment, the third member includes microlayer model container and microlayer model container upper cover, micro- liquid
Drop container is made of hard plastic material, and microlayer model container upper cover is made of soft plastic material；When fixed, the microlayer model holds
Device upper cover is cooperated with the detent slot, realizes that the first component and the third member are removably secured connection.
In one embodiment, it is provided in the detent slot from the puncture outstanding of the lower surface of the first component
Needle, the puncture needle are hollow；When the first component and fixed third member disassembly, the puncture needle penetrates micro- liquid
Drip container upper cover.
In one embodiment, the first component is provided with the sample-adding and inspection of at least one oil removal and upper oil slick
The collector unit of waste liquid after survey, preferably four, eight and 12；The second component is provided with the inspection of at least one microlayer model
Survey unit, preferably four, eight and 12；Third member is to include at least one equipped with microlayer model progress microlayer model PCR
The container of amplification, preferably four, eight and 12；Between each after the sample-adding and detection of oil removal and upper oil slick waste liquid receipts
Collect each the microlayer model detection unit and each appearance equipped with microlayer model progress microlayer model PCR amplification of unit corresponding thereto
Device cooperation, completes the detection of microlayer model after amplification.
In one embodiment, the collector unit of waste liquid includes after the sample-adding and detection of oil removal and upper oil slick between described
Above the first component upper surface setting upper oil slick loading slot, upper oil slick sample-adding through-hole, oil removal loading slot and
Oil removal is loaded through-hole, wherein floatings oil sample-adding through-hole and it is described between oil removal be loaded through-hole and be separately positioned on the upper oil slick and add
Sample slot and it is described between oil removal loading slot bottom；It is upper with being connected in place of the upper oil slick sample-adding through-hole of first component lower surface
Oil slick sample channel, described in the first component lower surface between connect in place of oil removal sample-adding through-hole between oil removal sample channel.
In one embodiment, the lower surface below the first component is provided with waste liquid tank, for collecting detection
All waste liquids afterwards.
In one embodiment, be provided with below the second component by by waste liquid tank residual air discharge
Structure.
In one embodiment, the upper oil slick loading slot and it is described between the volume of oil removal loading slot be respectively 1~900
Microlitre, preferably 5~500 microlitres, more preferably 100~200 microlitres.
In one embodiment, the microlayer model detection unit includes microlayer model signal piping, interval oil-in, interval
Oil pipe line, microlayer model container, microlayer model floating hole and microlayer model pipeline；Between oil removal from the interval oil-in enter the interval
Oil pipe line, microlayer model enter the microlayer model pipeline by microlayer model floating hole from the microlayer model container；The spacer tubing
Road and the microlayer model pipeline form criss-cross construction before the microlayer model signal piping, so that oil removal will be described micro-
Microlayer model separates in drop pipeline.
In one embodiment, described ten are formed by two spacer tubing roads and a microlayer model pipeline
Word intersection construction.
In one embodiment, hollow flow resistance area is provided in the pipeline after the interval oil-in；And/or it is described
An oil removal filtering area is provided in spacer tubing road.
In one embodiment, behind interval oil stream hollow flow resistance area excessively, it is divided into two-way and respectively enters described in two
Spacer tubing road.
In one embodiment, the microlayer model detection unit further includes floating oil-in, floating oil pipe line and floating
Oily connecting hole, upper oil slick are injected from the floating oil-in, by the floating oil pipe line, and from the upper oil slick connecting hole into
Drop receptacle in a subtle way squeezes the microlayer model in microlayer model container, so that microlayer model is floated by microlayer model floating hole
Into the microlayer model pipeline.
In one embodiment, oil slick filtering area is provided in the floating oil pipe line.
In one embodiment, positioning auxiliary channel is provided with beside the microlayer model signal piping.
In one embodiment, observation window is also set up on the second component, for cooperating optical system real-time monitoring
The microlayer model of generation.
In one embodiment, the microlayer model detection system further includes the 4th component, and gives described first by it
Oil removal and upper oil slick apply external pressure among component.
In one embodiment, it is also respectively provided on the first component and the second component convenient for the two fixation
Connect the location hole positioned.
Microlayer model detection system of the invention, can achieve following effect: (1) quick, reliable, parallel detection micron dimension
The fluorescence signal of " Water-In-Oil " microlayer model sample.(2) first component of the device and second component using thermoplastic material (such as
PC, COP, PMMA) processing, material and batch machining are low in cost, and (3) cooperate external pressure source, use the microlayer model detection system
System, the detection rates high speed of microlayer model sample are controllable.(4) integrated form microlayer model chip designs, and whole process is not likely to produce intersection
The EP pipe of pollution, the microlayer model sample after containing PCR is directly connect with chip, and without uncapping, the waste liquid after detection can be collected into
In waste collection pond, whole process is not likely to produce cross contamination.(5) microlayer model fluorescence detection chip operation is convenient.Only need hand
Oil removal and upper oil slick between dynamic pre-add, then cooperate optical detecting instrument, so that it may easily complete entire detection process.
Detailed description of the invention
It in order to more clearly explain the technical solutions in the embodiments of the present application, below will be to needed in the embodiment
Attached drawing is briefly described, it should be apparent that, the accompanying drawings in the following description is only some embodiments as described in this application, right
For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings
Its attached drawing.
Fig. 1 is the microlayer model detection system structure of one embodiment of the present invention；
Fig. 2 is the first component structural schematic diagram of one embodiment of the present invention；
Fig. 3 is that the first component of one embodiment of the present invention is illustrated by detent slot and third member connection structure
Figure；
Fig. 4 is the third member structural schematic diagram of one embodiment of the present invention；
Fig. 5 is the second component structural schematic diagram of one embodiment of the present invention；
Fig. 6 is oil removal hollow flow resistance area schematic diagram between one embodiment of the present invention；
Fig. 7 is the spacer tubing road filtering area schematic diagram of one embodiment of the present invention；
Fig. 8 is the upper oil slick driving microlayer model floating structural schematic diagram of one embodiment of the present invention；
The criss-cross construction schematic diagram of Fig. 9 one embodiment of the present invention；With
The signal piping structural schematic diagram of Figure 10 one embodiment of the present invention.
Specific embodiment
In order to make art technology field personnel more fully understand the technical solution in the application, below in conjunction with embodiment
The invention will be further described, it is clear that and described embodiments are only a part of embodiments of the present application, rather than whole
Embodiment.Based on the embodiment in the application, those of ordinary skill in the art are obtained without making creative work
The all other embodiment obtained, shall fall within the protection scope of the present application.The present invention is made with reference to the accompanying drawings and embodiments
It further describes.
One, microlayer model detection system structure
As shown in Figure 1, microlayer model generating means of the invention include: the first component 1, second component 2 and third member 3,
The first component 1 be the oil removal of microlayer model detection time and upper oil slick sample-adding and detection after waste collecting device, second component 2 is
Microlayer model detection system, third member 3 are the containers that microlayer model carries out microlayer model PCR amplification.Second component 2 is fixedly connected on
The upper surface of one component 1；Third member 3 is securely removably connected at the lower surface of the first component 1.
As shown in Figure 1, in some embodiments, microlayer model generating means of the invention further include the 4th component 4, the 4th
Component 4 is for sealing oil removal loading slot and upper oil slick loading slot between the first component 1.4th component 4 such as watertight potting pad,
Such as prepared with silica gel, which can be set stomata, and for applying pressure by it, effect is to guarantee outside
Crimping air-tightness between pressure source and the first component, and convenient for applying external pressure.
As shown in Figure 1, in some embodiments, processing location hole in the first component and/or second component 2, being convenient for
Cooperation each other.
As shown in Fig. 2, the first component 1 be realize the oil removal of microlayer model detection time and upper oil slick sample-adding and detection after give up
The collection of liquid.As shown in Fig. 2, in the present embodiment, between 8 are arranged in parallel on the first component 1 sample-adding of oil removal and upper oil slick with
And after detection waste liquid collector unit, from left to right, it is each above the upper surface of the first component 1 between oil removal and upper oil slick sample-adding
And after detection waste liquid the equidistant arranged in parallel of collector unit on the first component 1, the sample-adding of oil removal and upper oil slick and
Each collector unit of microlayer model is matched with microlayer model detection unit each on second component 2 after detection, is examined for microlayer model
Survey the microlayer model waste collection between unit after oil removal and the addition of oil sample sample and detection.The upper surface of 1 top of the first component
It is provided with oil slick loading slot 111, upper oil slick sample-adding through-hole 112, oil removal loading slot 113 and oil removal sample-adding through-hole 114,
In upper oil slick sample-adding through-hole 112 and an oil removal sample-adding through-hole 114 be separately positioned on oil slick loading slot 111 and oil removal loading slot
113 bottom centers；With in place of the upper oil slick of the 1 lower surface first component 1 of the first component is loaded through-hole 112 the upper oil slick that connects into
Sample pipeline 115, oil removal sample channel 116 between being connected in place of oil removal sample-adding through-hole 114 between the first component 1.
In some embodiments, upper oil slick sample channel 115 and an oil removal sample channel 116 are on the first component 1
The lower surface of side processes pipeline, then is packaged to form closed pipeline with case chip.
As shown in figure 3, in the lower surface of the first component 1, in upper oil slick sample channel 115 and oil removal sample channel 116
Lower position is provided with the detent slot 117 of third member 3, which is removably secured connection third member 3；?
It is provided in one component, 1 detent slot 117 from the lower surface of the first component 1 puncture needle 118 outstanding, which is hollow
, upper oil slick enters third member 3 by it.
Lower surface below the first component 1 is provided with waste liquid tank 119, will be glimmering for collecting all waste liquids after detecting
The microlayer model that light detection is crossed carries out hermetic reclaim.
As shown in figure 4, third member 3 includes setting on the microlayer model container 31 and the available first component of microlayer model for accommodating
The upper cover 32 that the puncture needle 118 set punctures, microlayer model container 31 is usually made of hard plastic material, for example, microlayer model container
31 be common centrifuge tube (EP pipe), and upper cover 32 is usually made of soft plastic material, convenient for being punctured with puncture needle 118, such as
Upper cover 32 is the centrifuge tube lid made of organic silicon rubber.In some embodiments, each microlayer model container 31 and component 1
The collector unit of waste liquid matches after the sample-adding and detection of oil removal and upper oil slick between each.In some embodiments, each
Microlayer model container 31 links together.In the present embodiment, 8 microlayer model containers 31 link together, and 8 upper covers 32 connect
Together；They are integrally machined respectively.
As shown in figure 5, in the standard optical disc that outside diameter is 118mm, interior circular diameter is 22mm, from left to right, one
It equidistantly arranges in a standard optical disc 8 identical microlayer model detection units 21, is used for parallel fluorescence detection microlayer model.
The heart has a centre bore 22 in the chips, the centre bore 22 from CD processing technology, for inject injection molding material with
And during mass production substrate transhipment, and in mass fluorescence detection microlayer model detection system turn
Fortune.Typical round compact disk structure is not easy to position, and is eight-sided formation by chip manufacture, and process two location holes 23, is convenient for
The location fit of microlayer model chip and relevant device.In the respectively equidistantly identical microlayer models detection of arrangement 4 of centre bore two sides
Unit 21 is used for parallel detection microlayer model.
As shown in figure 5, each drop detection unit 1 includes: interval oil-in 2111, hollow flow resistance area from top to bottom
2112, oil removal filtering area 2114, the upper oil slick filtering area in floating oil-in 2121, one between two 2113, two, spacer tubing roads
2122, floating oil pipe line 2123 and upper oil slick connecting hole 2124, microlayer model floating hole 213, microlayer model pipeline 214, microlayer model
Signal piping 215 and microlayer model waste liquid port 2126；Upper oil slick connecting hole 2124 is connection floating oil pipe line 2123 and third member 3
The through-hole of (in lower section, being not shown).Microlayer model detection system as shown in Figure 1 repairs the normal structure of general disc
Just, the space of CD, arranged in parallel microlayer model detection channel can be utilized to greatest extent.Meanwhile it being processed using precise injection molding technique
Chip, designed in conjunction with hollow flow resistance area and filtering area, quickly, the reliable micron dimension " Water-In-Oil " uniform using fluorescence detection
Microlayer model.
In one embodiment, in 8 drop detection units 1 of Fig. 5, it is each interval oil-in 111 between away from
From the distance between the distance between, floating oil-in 121 microlayer model floating hole 13, be it is equal, which is equal to standard
The distance between eight channel pipettor suction nozzles.
As shown in fig. 6, an oil removal is injected into interval oil-in 2111 first with external air pump or peristaltic pump.For
Accurate control room oil removal phase sample injection volume is provided with hollow flow resistance area 2112, accurate control room in one embodiment
Oil removal sample volume.Between oil removal can infiltrate in surface of polymer material, in the case where not applying pressure condition, by capillarity, interval
Oil automatically flows into microchannel.Under extreme case, oil removal under capillary action, constant flow.Design back-shaped flow resistance area 2112
Purpose be accurate control room oil removal sample volume, reduce constant flow of the oil removal under capillary action in microchannel to the greatest extent,
So that oil removal sample volume is only controlled by external air pump or peristaltic pump.
Then, oil removal is by an oily phase split entry, the spacer tubing road into same design being divided into two
In 2113, effect is that an oil removal crosses at right-angled intersection with floating drop, pushes microlayer model movement；And it is imitated by sheath stream
It answers, floating drop is expressed to runner center, convenient for the fluorescence signal in detection microlayer model.As shown in fig. 7, oil removal is each between two-way
Oil removal filtering area 2114 between into one, filtering area 2114 is one group of columnar arrays structure, as shown in fig. 7, columnar arrays structure
There is multiple rows of columnar arrays staggered.Between impurity present in oil removal (particle, silk wadding fiber etc.) be blocked in this group of column knot
At structure, influence of the impurity for drop fluorescence detection is eliminated.
As shown in figure 8, upper oil slick flows into floating oil pipe line 2123 under outer boundary's gas pressure, from floating oil-in 2121,
Under being flowed at upper oil slick connecting hole 2124, flow into third member 3 (in lower section, being not shown), while micro- in third member 3
Drop floats under the effect of upper oil slick from microlayer model floating hole 13.Third member 3 is placed on oil slick connecting hole 2124 and micro- liquid
It drips below floating hole 213, the line space design in upper oil slick connecting hole 2124 and microlayer model floating hole 213 is with reference to third member 3
Size, if third member 3 is EP pipe, the spacing between upper oil slick connecting hole 2124 and microlayer model floating hole 213 is less than EP
The width of pipe.
As shown in figure 9, two spacer tubing roads 2113 and a microlayer model pipeline 214 form criss-cross construction, purpose
It is that the compact arranged microlayer model for carrying out floating is spaced apart by oil removal between two sides, and makes the signal interference between microlayer model
It reduces；Microlayer model single layer row can be controlled by the air pressure of oil removal between control two sides by " criss-cross construction " design simultaneously
The distance of cloth.In the microlayer model signal piping after " cross structure ", after microlayer model is spaced apart, the stream of flowing in one line
Cross carry out optical detection.
As shown in Figure 10 after " criss-cross construction ", microlayer model is single equidistant in microlayer model signal piping at this time
Arrangement is come, while the position of constant spacing is provided with a closing assist gallery by microlayer model signal piping to be detected
2151.Without oil phase, water phase, microlayer model sample flow in closed channel, the regional imaging, light transmission/astigmatism property are stablized, just
215 center of microlayer model signal piping is navigated in light path detection.
After the detection of a large amount of microlayer model, the microlayer model that will test signal is flowed out from microlayer model waste liquid port 216, is flowed into
In the closing collecting tank entrance of one outside.The closed purpose of collecting tank is cross contamination caused by preventing microlayer model waste liquid.Such as
Shown in Fig. 5, in some embodiments, microlayer model detection system is provided with exhaust entrance 2171, the gas exhaust piping of collecting tank
2172 and air exit 2173, exhaust entrance 2171 and the outlet of closing collecting tank connect, and by gas exhaust piping 2172 and are vented out
Mouth 2173 connects, in order to discharge the pressure for continuing to flow into liquid generation in collecting tank, while reduce cross contamination.
Two, microlayer model detection process
The microlayer model detection course of work of the invention is as follows.Firstly, the microlayer model generated is collected into microlayer model container 31
In, for example, EP pipe.Then, closed EP pipe upper cover 32 is assembled on EP pipe, is put into Standard PCR instrument and is carried out pcr amplification reaction.
Then the above-mentioned third member equipped with amplification sample is subjected to assembly connection by detent slot 117 and the first component 1, connected in assembly
Puncture needle 118 completes the puncture to upper cover 32 in termination process.
Upper oil slick is loaded in advance in the upper oil slick loading slot 111 of the first component 1.Under external pressure effect, float
Oil flows into upper oil slick sample channel 115 from sample-adding through-hole 112, and then upper oil slick enters upper oil slick from floating oil-in 2121 and filters
Area 2122, floating oil pipe line 2123 and upper oil slick connecting hole 2124, then by puncture needle 118, into microlayer model container 31
In.Since the density of upper oil slick is greater than microlayer model density in microlayer model container 31, microlayer model is constantly in ullage.
Then floating oil pipe line 1222 enters microlayer model container 31 by the middle vacancy of floating oilhole 224 and puncture needle 118
In.32 airtight connection of upper cover of the first component 1 and third member 3, floating oilhole 224, which is arranged in, to be corresponded in puncture needle 118
Vacancy；Upper oil slick can only flow into third member 31 from the middle vacancy of puncture needle 118.Since the density of upper oil slick is greater than third portion
Microlayer model density, microlayer model are constantly in ullage in part 331.It is upper passing through during puncture needle 118 punctures upper cover 32
118 surrounding of puncture needle of lid 32 forms gap, and for the microlayer model in third member 3 under the action of upper oil slick, floating passes through puncture
Gap around needle 118, since the first component 1 is closely connect with second component 2, the drop of floating is merely able to enter second component
Microlayer model floating hole 213 on 2, subsequently into microlayer model pipeline microlayer model pipeline 214.
Between oil removal by an oily phase split entry, in the spacer tubing road 2113 into same design being divided into two,
Its effect is that an oil removal crosses at right-angled intersection with floating drop, pushes microlayer model movement；And by sheath stream effect, will float
Drop is expressed to runner center, convenient for the fluorescence signal in detection microlayer model.Oil removal is filtered between oil removal respectively enters one between two-way
Area 2114, filtering area 2114 are one group of columnar arrays structures, and columnar arrays structure has multiple rows of columnar arrays staggered.Between oil removal
Present in impurity (particle, silk wadding fiber etc.) be blocked at this group of column structure, eliminate impurity for drop fluorescence detection
Influence.
Two spacer tubing roads 2113 and a microlayer model pipeline 214 form criss-cross construction, it is therefore an objective to make to float and
Compact arranged microlayer model be spaced apart by oil removal between two sides, and the signal interference between microlayer model is reduced；Simultaneously
The spacing of microlayer model single layer arrangement can be controlled by the air pressure of oil removal between control two sides by " criss-cross construction " design
Distance.In the microlayer model signal piping after " cross structure ", after microlayer model is spaced apart, carry out light is flowed through in flowing in one line
Learn detection.
After " criss-cross construction ", microlayer model comes in the single equidistant arrangement of microlayer model signal piping at this time,
The position of constant spacing is provided with a closing assist gallery 2151 by microlayer model signal piping to be detected simultaneously.It is closed
Without oil phase, water phase, microlayer model sample flow in channel, the regional imaging, light transmission/astigmatism property are stablized, are detected convenient for optical path
System navigates to 215 center of microlayer model signal piping.
After the detection of a large amount of microlayer model, the microlayer model that will test signal is flowed out from microlayer model waste liquid port 216, is flowed into
In the closing collecting tank entrance of one outside.The closed purpose of collecting tank is cross contamination caused by preventing microlayer model waste liquid.
It should be understood that the present invention disclosed is not limited only to specific method, scheme and the substance of description, because these
It is alterable.It will also be understood that purpose of the terminology used here just for the sake of the specific embodiment scheme of description, rather than
It is intended to limit the scope of the invention, the scope of the present invention is limited solely by the attached claims.
Those skilled in the art, which will also be appreciated that or be able to confirm that, uses no more than routine experiment, institute herein
The many equivalents for the specific embodiment of the invention stated.These equivalents are also contained in the attached claims.
Claims (22)
1. a kind of microlayer model detection system, it is characterised in that:
The microlayer model detection system includes the first component, second component and third member；
The dress when first component is microlayer model detection for waste collection after the sample-adding and detection of oil removal and upper oil slick
It sets；
The second component is microlayer model detection chip, and the microlayer model detection system includes a centre bore, the centre bore
For being injected in the microlayer model detection system preparation process during injection molding material and mass production and/or mass fluorescence
The transhipment of microlayer model detection system in detection process；
The container that microlayer model carries out microlayer model PCR amplification is equipped with the third member；
The second component is fixedly connected on the upper surface of the first component；Connection is removably secured with the third member
In the lower surface of the first component.
2. microlayer model detection system according to claim 1, it is characterised in that: the first component and the second component
It is sealedly and fixedly connected by dispensing mode or ultrasonic welding mode.
3. microlayer model detection system according to claim 1, it is characterised in that: the first component and the second component
It is formed respectively by integrated injection molding.
4. microlayer model detection system according to claim 1, it is characterised in that: the first component and the second component
It is thermoplastic material, preferably makrolon material, cyclic olefine copolymer or polymethyl methacrylate, polypropylene.
5. microlayer model detection system according to claim 1, it is characterised in that: the first component is provided with for fixing
The detent slot of the third member.
6. microlayer model detection system according to claim 5, it is characterised in that: the third member includes microlayer model container
With microlayer model container upper cover, the microlayer model container is made of hard plastic material, and microlayer model container upper cover is by soft plastic material
Material is made；When fixed, the microlayer model container upper cover is cooperated with the detent slot, realizes the first component and described the
Three components are removably secured connection.
7. microlayer model detection system according to claim 6, it is characterised in that: be provided in the detent slot from described
The lower surface of first component puncture needle outstanding, the puncture needle are hollow；The first component and third member disassembly are solid
Periodically, the puncture needle penetrates the microlayer model container upper cover.
8. microlayer model detection system according to claim 1, it is characterised in that:
The first component is provided with the sample-adding of at least one oil removal and upper oil slick and the collector unit of waste liquid after detection, excellent
It is selected as four, eight and 12；
The second component is provided at least one microlayer model detection unit, preferably four, eight and 12；
The container for microlayer model PCR amplification that third member is that include at least one carry out equipped with microlayer model, preferably four, eight and
12；
Each microlayer model of the collector unit of waste liquid corresponding thereto after the sample-adding and detection of oil removal and upper oil slick between each
Detection unit and each container for carrying out microlayer model PCR amplification equipped with microlayer model cooperate, and complete the detection of microlayer model after amplification.
9. microlayer model detection system according to claim 8, it is characterised in that: between described the sample-adding of oil removal and upper oil slick with
And the collector unit of waste liquid includes upper oil slick loading slot, the upper oil slick of the upper surface setting above the first component after detection
Be loaded through-hole, oil removal loading slot and oil removal and be loaded through-hole, wherein floatings oil sample-adding through-hole and it is described between oil removal sample-adding
Through-hole be separately positioned on the upper oil slick loading slot and it is described between oil removal loading slot bottom；With described in the first component lower surface
The upper oil slick sample channel connected in place of upper oil slick sample-adding through-hole, described in the first component lower surface between oil removal sample-adding it is logical
Oil removal sample channel between connection in place of hole.
10. microlayer model detection system according to claim 9, it is characterised in that: the following table below the first component
Face is provided with waste liquid tank, for collecting all waste liquids after detecting.
11. microlayer model detection system according to claim 10, it is characterised in that: be provided with below the second component
The structure that residual air in waste liquid tank will be discharged.
12. microlayer model detection system according to claim 9, it is characterised in that: the upper oil slick loading slot and it is described between
The volume of oil removal loading slot is respectively 1~900 microlitre, preferably 5~500 microlitres, more preferably 100~200 microlitres.
13. microlayer model detection system according to claim 8, it is characterised in that: the microlayer model detection unit includes micro-
Drop detection pipeline, interval oil-in, spacer tubing road, microlayer model container, microlayer model floating hole and microlayer model pipeline；Between oil removal
Enter the spacer tubing road from the interval oil-in, microlayer model is entered from the microlayer model container by microlayer model floating hole
The microlayer model pipeline；The spacer tubing road and the microlayer model pipeline form cross before the microlayer model signal piping
Intersection construction, so that oil removal separates microlayer model in the microlayer model pipeline.
14. microlayer model detection system according to claim 13, it is characterised in that: by two spacer tubing roads and
One microlayer model pipeline forms the criss-cross construction.
15. microlayer model detection system according to claim 13, it is characterised in that: the pipeline after the interval oil-in
In be provided with hollow flow resistance area；And/or an oil removal filtering area is provided in the spacer tubing road.
16. microlayer model detection system according to claim 15, it is characterised in that: interval oil stream hollow flow resistance area excessively
Afterwards, it is divided into two-way and respectively enters two spacer tubing roads.
17. microlayer model detection system according to claim 13, it is characterised in that: the microlayer model detection unit further includes
Floating oil-in, floating oil pipe line and upper oil slick connecting hole, upper oil slick are injected from the floating oil-in, pass through the upper oil slick
Pipeline, and enter microlayer model container from the upper oil slick connecting hole, the microlayer model in microlayer model container is squeezed, so that micro- liquid
Drop is floated by microlayer model floating hole enters the microlayer model pipeline.
18. microlayer model detection system according to claim 17, it is characterised in that: be provided in the floating oil pipe line
Oil slick filtering area.
19. microlayer model detection system according to claim 13, it is characterised in that: beside the microlayer model signal piping
It is provided with positioning auxiliary channel.
20. -19 any microlayer model detection system according to claim 1, it is characterised in that: also set up on the second component
Observation window, the microlayer model for cooperating optical system real-time monitoring to generate.
21. -19 any microlayer model detection system according to claim 1, it is characterised in that: the microlayer model detection system is also
Apply external pressure to oil removal among the first component and upper oil slick including the 4th component, and by it.
22. -19 any microlayer model detection system according to claim 1, it is characterised in that: the first component and described
It is also respectively provided on two components and is fixedly connected with the location hole positioned convenient for the two.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711074989.0A CN109746063B (en) | 2017-11-06 | 2017-11-06 | Micro-droplet detection system |
| EP18872941.2A EP3674393A4 (en) | 2017-11-06 | 2018-11-03 | Droplet detection apparatus |
| PCT/CN2018/113852 WO2019086019A1 (en) | 2017-11-06 | 2018-11-03 | Droplet detection apparatus |
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| CN201711074989.0A CN109746063B (en) | 2017-11-06 | 2017-11-06 | Micro-droplet detection system |
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| CN109746063B CN109746063B (en) | 2024-05-24 |
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