CN107020164B - A kind of high throughput micro particles circulation sorting is with enrichment facility and preparation method thereof - Google Patents
A kind of high throughput micro particles circulation sorting is with enrichment facility and preparation method thereof Download PDFInfo
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- CN107020164B CN107020164B CN201710235635.3A CN201710235635A CN107020164B CN 107020164 B CN107020164 B CN 107020164B CN 201710235635 A CN201710235635 A CN 201710235635A CN 107020164 B CN107020164 B CN 107020164B
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- 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
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- 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
- B01L3/502715—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 interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
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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
- B01L3/50273—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 the means or forces applied to move the fluids
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- 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
- B01L3/502738—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 integrated valves
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- 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
- B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
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- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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Abstract
The present invention discloses a kind of high-throughput micro particles circulation sorting and enrichment facility and preparation method thereof, the device is sequentially connected with inner outlet solution receiving flask (8), check valve (9), peristaltic pump (1), flow dispensing connector (2), multiple-layer stacked micro-fluidic chip (3), there is external solution outlet (16) and interior liquid outlet (17) on multiple-layer stacked micro-fluidic chip (3), it is separately connected outer vent solution receiving flask (7) and inner outlet solution receiving flask (8), and is connected between each component by connecting hose (10~14)；Making step is as follows: 1) making multiple-layer stacked micro-fluidic chip (3)；2) each component is sequentially connected with by connecting hose (10~14)；This method is simple, sorting is stablized, flux is big, thickening efficiency is high, can realize that the sorting to micron order biomone is concentrated, and can be widely used for the environment measuring of the medical diagnosis and algae bio of haemocyte etc. using the microfluid inertia effect.
Description
Technical field
The present invention relates to a kind of high-throughput micro particles circulation sortings and enrichment facility and preparation method thereof, belong to micro-fluidic
Chip field.
Background technique
Nowadays particle sorting and concentration technique it is nearly ubiquitous, and often including biological study, chemical analysis,
Important link including environment measuring and medical diagnosis in many industries.Due to micro-fluidic chip collection it is small in size, high-throughput, low at
This and it is easy of integration the advantages that, so as to so that it can be adapted for various business and industry purpose, also because
This obtains the concern of many researchers.Particle sorting at present and method for concentration have centrifugal process, flow cytometry, chromatography
Method, Fluorimetric screening test etc..
The difference that centrifugal process utilizes different particles to settle in centrifugal force field, under same centrifugal condition, sinking speed
Difference makes the size in a non-uniform mixed liquid, variform particle branch sink by being continuously increased relative centrifugal force
It forms sediment.This method resolution ratio is not high, and various particles of the factor in precipitation in the same order of magnitude are not easy to separate, and is usually used in other separation
Semifinished product before means extracts, and cell sample may be destroyed in centrifugal process.
Flow cytometry is that the single cell suspension through fluorescent staining or label is full of in sheath fluid flow chamber with high pressure indentation.?
Under the package and promotion of sheath fluid, cell be ordered in it is single-row, with certain speed from flow chamber spout spray.The liquid stream of ejection is in superelevation
It is fractured into uniform drop under the piezo-electric crystal effect of frequency, these drops are filled with positive and negative different charge, when drop stream passes through
When with several kilovolts of deflecting plates, deflects, fallen into respective collection vessel under the action of high voltage electric field, the liquid not charged
It is dropped into intermediate waste fluid container, to realize the separation of cell.Though this method has good sorting index, disadvantage is also brighter
It is aobvious, it can only complete to operate in laboratory, and apparatus structure is complicated, needs to complete the sorting of sample by electrical equipment.
Red, orange, green, blue, yellow (ROGBY) is to be attracted the solute phase in stationary phase more by force using particle in the two-phase performed relative motion
To lagging behind the solute attracted in mobile phase strongly, it is repeated and repeated dispensing, makes in mixture with mobile
Each component is separated.This method has the advantages that separative efficiency is high, separating rate is fast, but its qualitative research ability is poor, and instrument
Device equipment is expensive.
Fluorimetric screening test is that have according to material molecule Absorption and fluorescence spectrum energy level transition mechanism and absorb photon energy
The substance of power can launch the principle of light longer than excitation wavelength in moment under special wavelength light (such as ultraviolet light) irradiation
Complete particle sorting.This method has the advantages that sensitivity for analysis is high, selectivity is strong and uses object that is easy, but itself can fluorescing
Matter is relatively fewer, and disturbing factor is more extremely stringent for experimental situation requirement.
Inertia lift that inertia microflow control technique is subject to using the inertia migration effect under particle minute yardstick and in curved stream
The Secondary Flow that is generated in road and (two rotations are generated in vertical main flow direction when fluid is by curved runner by Dean drag
Turn contrary vortex, referred to as Dean stream, due to Dean stream introducing and generate Dean drag) collective effect realizes to micro-
The accurate control of grain becomes the new tool of particle sorting concentration.Micro-fluidic chip has required sample solution small, analysis efficiency
Height, be easy to encapsulate so as to realize high-throughput detection, system integration, micromation, automation and it is portable etc. obviously
The advantages of.But current technology mainly uses single layer normal spiral runner chip, and particle is unstable under low discharge, and needed for it
Sorting flow is higher to be required also to correspondingly increase to chip bonding, and the analysis time used is unable to complete instant detection.And in order to improve
Flow reduces the processing time and the multi-layer micro-fluidic chips that make mostly use hole knockout from top to down at present, there is each layer
The non-uniform problem of chip flow.
Summary of the invention
Technical problem: the object of the present invention is to provide a kind of high-throughput micro particles sortings of circulation and enrichment facility and its system
Make method.The micro fluidic device structure and manufacture craft are simple, low in cost, sorting thickening efficiency is high, are applicable not only to test
Room scientific research is equally applicable for biomedical diagnostic, environment and the commercial objects such as detects immediately.
Technical solution: the present invention provides a kind of high-throughput micro particles circulation sortings and enrichment facility, the device to include
Peristaltic pump, flow dispensing connector, multiple-layer stacked micro-fluidic chip, outer vent solution receiving flask, inner outlet solution receiving flask and list
To valve and connection hose, in which:
The inner outlet solution receiving flask is also initial sample when micro particles circulation sorting starts with concentration process simultaneously
The placement bottle of liquid；
Described check valve one end connects inner outlet solution receiving flask by connection hose, and the other end passes through connection hose connection
The import of peristaltic pump, liquid can only flow to peristaltic pump by inner outlet solution receiving flask；
One end of the flow dispensing connector has macropore, and the other end has aperture, and the aperture passes through connection hose
It is connected with the outlet of peristaltic pump, the side inlet opening of the macropore and multiple-layer stacked micro-fluidic chip is connected directly；
The multiple-layer stacked micro-fluidic chip is from top to bottom by top layer chip, middle layer chip and basal layer chip bonding group
At, wherein at least one layer of middle layer chip, contact surface position, the adjacent middle layer core of the top layer chip and middle layer chip
It is nested that the contact surface position of piece and middle layer chip with the corresponding position at the contact surface position of basal layer chip contain spiral
Sinusoidal runner and side inlet opening, wherein the liquid feeding end of the nested sinusoidal runner of spiral is located at the outmost turns of helical structure, and and side
Face inlet opening communicates, and the nested sinusoidal runner outlet end of spiral is located at the innermost circle of helical structure, goes out in the nested sinusoidal runner of spiral
The interior direction at liquid end is provided with interior fluid hole, and the interior fluid hole is communicated with the outlet end of the nested sinusoidal runner of spiral, and is passed through
Connection hose is connected with inner outlet solution receiving flask, is provided with outer liquid outlet holes in the lateral direction of the nested sinusoidal runner outlet end of spiral
Hole, the outgoing fluid apertures is communicated with the outlet end of the nested sinusoidal runner of spiral, and is collected by connection hose and outer vent solution
Bottle is connected.
Wherein:
The peristaltic pump, flow dispensing connector, multiple-layer stacked micro-fluidic chip, outer vent solution receiving flask, inner outlet
Connection between solution receiving flask, check valve and connection hose is to be tightly connected.
The outer vent solution receiving flask is waste collection bottle.
The lower surface of the middle layer chip have the nested sinusoidal flow passage structure of concave spiral and concave side into
Fluid apertures structure, upper surface are burnishing surface；The structure of the top layer chip is identical with the structure of middle layer chip, and its thickness is greater than
The thickness of middle layer chip；Two surfaces up and down of the basal layer chip are burnishing surface.
The shape of the nested sinusoidal runner of the spiral is to be superimposed sine curve on helical curve, by the concave spiral
The burnishing surface of nested sine flow passage structure and adjacent core lamella is bonded, and the section of the nested sinusoidal runner of the spiral is square
Shape, width of flow path are 500-700 μm, and runner height is 50 μm -200 μm.
The corresponding non-helical nested sine runner of the top layer chip and middle layer chip and non-side feed liquor hole site
Place is provided with location hole, and the quantity of the location hole is 2 or 4；The high 5-10mm of top layer chip, middle layer
The high 2-5mm of chip.
The basal layer chip material is in dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate
One kind, top layer chip material is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate,
Middle layer chip material is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate.
Described check valve one end by connection hose connects connection hose in inner outlet solution receiving flask, and to protrude into inner outlet molten
There are following relational expressions between the depth and micro particles cocnentration factor of liquid receiving flask:
Wherein n is micro particles cocnentration factor, and V is the volume of sample liquid after concentration, and U is the volume of initial sample liquid.
The present invention also provides the production method of a kind of high-throughput micro particles circulation sorting and enrichment facility, this method packets
Include following steps:
1) multiple-layer stacked micro-fluidic chip is made:
1. preparing top layer chip, middle layer chip and basal layer chip；
2. top layer chip, middle layer chip are accurately bonded by location hole, so that the nested sinusoidal runner of each layer of spiral
Upper and lower overlapping alignment opens interior fluid hole, outer on the inside of each layer of the nested sinusoidal runner outlet end of spiral from top to bottom later
Fluid hole over side, obtains bonding structure；
3. bonding structure is finally bonded to basal layer chip upper surface, it is put into baking oven heating later, obtains multiple-layer stacked
Micro-fluidic chip；
2) side inlet opening is opened in multiple-layer stacked micro-fluidic chip side, makes the liquid feeding end of its sinusoidal runner nested with spiral
It communicates, and is tightly connected with the macropore of flow dispensing connector.
3) it is sequentially connected each component using connection hose, constitutes the high-throughput micro particles circulation sorting and is filled with concentration
It sets.
Wherein:
1. specific step is as follows for prepare top layer chip, middle layer chip and the basal layer chip for the step: first
First, in the silicon wafer surface spin coating negative photoresist of cleaned drying and processing, and the photoetching technique system of printing film exposure mask is utilized
Make the formpiston of top layer chip, middle layer chip and basal layer chip；Secondly, top layer chip, middle layer chip and substrate will be prepared
The material of layer chip is poured on the anode membrane prepared, is put into baking oven heating；Finally, demoulding obtains top layer chip, middle layer core
Piece and basal layer chip.
The utility model has the advantages that compared with prior art, the invention has the following advantages that
1, micro fluidic device structure of the invention and manufacture craft are simple, low in cost；
2, the nested sinusoidal flow passage structure of spiral of the invention reduces the flow needed for die grading focuses, and increasing chip is having
Limit the safety and stability under bond strength；And space is more saved compared to traditional normal spiral runner and is conducive to be superimposed core
The micromation of piece；The flow dispensing connector of lateral opening hole helps to improve the flux of the sorting concentration of chip superposed；
3, the efficiency of sorting concentration is substantially increased, by the circular flow that electromagnetism check valve completes sorting concentration for sorting
Enriching service saves the plenty of time, can move towards detection immediately by laboratory testing to technology and have great significance, not only fit
It is studied for laboratory science, is equally applicable for biomedical diagnostic, environment and the commercial objects such as detects immediately.
Detailed description of the invention
Fig. 1 is the sorting of high-throughput micro particles circulation and enrichment facility structural schematic diagram of the invention；
Fig. 2 is the nested sinusoidal flow passage structure schematic diagram of spiral；
Fig. 3 is flow dispensing connector and multiple-layer stacked chip assembling schematic diagram；
Fig. 4 is chip inlet and exit particle distribution schematic diagram；
Fig. 5 is that fluid channel particle sorts concentration principle schematic diagram；
Fig. 6 is to sort Platymonas helgolandica var cell experiment effect picture using device；
Have in figure: peristaltic pump 1, flow dispensing connector 2, multiple-layer stacked micro-fluidic chip 3, top layer chip 4, middle layer chip
5, basal layer chip 6, outer vent solution receiving flask 7, inner outlet solution receiving flask 8, check valve 9, connection hose 10~14, spiral
Nested sine runner 15, outgoing fluid apertures 16, interior fluid hole 17, side inlet 18, location hole 19, aperture 20, macropore 21, particle
22, the parabola shaped velocity profile 23 of primary howl leaf stream, Dean flow 24, fluid drag force F1, shear-induced inertia lift F2, wall surface
Induce inertia lift F3, Dean drag F4。
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is further explained, but it is of the invention
Content is not limited solely to the following examples.
Working principle:
When the present invention recycles high-throughput micro particles sorting with enrichment facility sorting concentrating sample, it is placed on inner outlet solution receipts
Particle suspension in collection bottle 8 is imported into flow dispensing connector 2 by peristaltic pump 1 with specific flow velocity；Sample is matched through flow
Each layer multi-layer is entered with identical flow velocity to connector 2 and is superimposed micro-fluidic chip 3；Micro particles 22 in particle suspension are in core
It will be by fluid drag force F in piece1Effect and moved along flow direction；Due to the parabola shaped speed of howl leaf stream primary in runner
Section 23 is spent, micro particles will be by the shear-induced inertia lift F for being directed toward wall surface2；When micro particles lean near wall, due to
Micro particles are because the symmetrical tail that rotation rotation generates is generated wall surface induction inertia lift F by Wall effect3；When fluid passes through
The opposite vortex in two direction of rotation is generated when crossing curved runner in vertical main flow direction, referred to as Dean flows 24, due to
Dean stream introducing and generate Dean drag F4；The net inertia lift that particle is not only generated by inertia migration effect in curved runner
It influences and migrates, also by Dean drag F caused by Secondary Flow4It influences；Therefore, micro particles can be approximately considered to drag in Dean
It is migrated under the effect of the coupled in common of power and inertia lift to the side close to helical flow path inner wall；Most particulate particles flow into
Inner outlet solution receiving flask 8 then passes through check valve 9 and is again introduced into next circulation that peristaltic pump 1 carries out sorting concentration；Pass through
Hose protrudes into the cocnentration factor of the depth control apparatus processing sample solution of inner outlet solution receiving flask 8, protrudes into inner outlet solution receipts
There are following relational expressions between the depth and cocnentration factor of collection bottle 8:
Wherein n is micro particles cocnentration factor, and V is the volume of sample liquid after concentration, and U is the volume of initial sample liquid.
Above-mentioned net inertia lift are as follows: wall surface induces inertia lift F3With shear-induced inertia lift F2Resultant force, the active force
There are following relational expressions:
In formula, ρ is fluid density, UmFor maximum flow rate, apFor particle diameter, CLFor dimensionless number lift coefficient, DhFor stream
Road water conservancy diameter.
Above-mentioned Dean drag F4Are as follows: two direction of rotation are generated in vertical main flow direction when fluid passes through curved runner
Opposite vortex, referred to as Dean stream 24 generates Dean drag F since Dean flows 24 introducing4.Generally assumed that in research for
Its size of Stokes resistance estimate.There are following relational expressions for the active force:
FD=5.4 × 10-4πμDe1.63ap
μ is flow velocity, a in formulapFor particle diameter, DeFor Dean number, relational expression is as follows:
R in formulaeFor dimensionless number Reynolds number, R is runner radius of curvature.
It is the peristaltic pump 1, flow dispensing connector 2, multiple-layer stacked micro-fluidic chip 3, outer vent solution receiving flask 7, interior
Connection between outlet solution receiving flask 8, check valve 9 and connection hose 10~14 is to be tightly connected, and prevents suspension from leaking.
As shown in figures 1 to 6, the present invention provides a kind of high-throughput micro particles circulation sorting and enrichment facility, the device packets
Peristaltic pump 1, flow dispensing connector 2, multiple-layer stacked micro-fluidic chip 3, outer vent solution receiving flask 7, inner outlet solution is included to collect
Bottle 8 and check valve 9 and connection hose 10~14, in which:
The inner outlet solution receiving flask 8 is also initial sample when micro particles circulation sorting starts with concentration process simultaneously
The placement bottle of liquid；
Described 9 one end of check valve connects inner outlet solution receiving flask 8 by connection hose 13, and the other end passes through connection hose
The import of 14 connection peristaltic pumps 1, liquid can only flow to peristaltic pump 1 by inner outlet solution receiving flask 8；
One end of the flow dispensing connector 2 has macropore 21, and the other end has aperture 20, and the aperture 20 passes through company
It connects hose 10 to be connected with the outlet of peristaltic pump 1, the side inlet opening 18 of the macropore 21 and multiple-layer stacked micro-fluidic chip 3 is direct
It is connected；
The multiple-layer stacked micro-fluidic chip 3 is from top to bottom by 6 key of top layer chip 4, middle layer chip 5 and basal layer chip
It is combined into, wherein at least one layer of middle layer chip 5, it is the contact surface position of the top layer chip 4 and middle layer chip 5, adjacent
The corresponding position at the contact surface position of middle layer chip 5 and the contact surface position of middle layer chip 5 and basal layer chip 6 contains
There are the nested sinusoidal runner 15 of spiral and side inlet opening 18, wherein the liquid feeding end of the nested sinusoidal runner 15 of spiral is located at helical structure
Outmost turns, and communicated with side inlet opening 18, nested sinusoidal 15 outlet end of runner of spiral is located at the innermost circle of helical structure,
The interior direction of spiral 15 outlet end of nested sinusoidal runner is provided with interior fluid hole 17, and the interior fluid hole 17 is nested with spiral sinusoidal
The outlet end of runner 15 communicates, and is connected by connecting hose 12 with inner outlet solution receiving flask 8, in the nested sinusoidal runner of spiral
The lateral direction of 15 outlet ends is provided with outgoing fluid apertures 16, the outlet end phase of the outgoing fluid apertures 16 sinusoidal runner 15 nested with spiral
It is logical, and be connected by connecting hose 11 with outer vent solution receiving flask 7.
The multiple-layer stacked micro-fluidic chip 3 is from top to bottom by 6 key of top layer chip 4, middle layer chip 5 and basal layer chip
It is combined into, wherein at least one layer of middle layer chip 5, it is the contact surface position of the top layer chip 4 and middle layer chip 5, adjacent
The corresponding position at the contact surface position of middle layer chip 5 and the contact surface position of middle layer chip 5 and basal layer chip 6 contains
There is the nested sinusoidal runner 15 of spiral, the liquid feeding end of the nested sinusoidal runner 15 of spiral is communicated with side inlet opening 18, in spiral nesting
It is provided with interior fluid hole 17 on the inside of sinusoidal 15 outlet end of runner and is connected by connecting hose 12 with inner outlet solution receiving flask 8,
Outgoing fluid apertures 16 is provided on the outside of spiral 15 outlet end of nested sinusoidal runner and is received by connection hose 11 and outer vent solution
Collect bottle 7 to be connected；
Wherein:
It is the peristaltic pump 1, flow dispensing connector 2, multiple-layer stacked micro-fluidic chip 3, outer vent solution receiving flask 7, interior
Outlet solution receiving flask 8, check valve 9 and connection hose (10~14) are to be tightly connected.
The lower surface of the middle layer chip 5 has nested sinusoidal 15 structure of runner of concave spiral and concave side
18 structure of inlet opening, upper surface are burnishing surface；The structure of the top layer chip 4 is identical with the structure of middle layer chip 5, and it is thick
Degree is greater than the thickness of middle layer chip 5；Two surfaces up and down of the basal layer chip 6 are burnishing surface.
The shape of the nested sinusoidal runner 15 of the spiral is to be superimposed sine curve on helical curve, by the concave spiral shell
Nested sinusoidal 15 structure of runner of rotation and the burnishing surface of adjacent core lamella are bonded, the cross section of fluid channel of the nested sinusoidal runner 15 of spiral
For rectangle, width of flow path is 500-700 μm, and runner height is 50 μm -200 μm.
The corresponding non-helical nested sine runner 15 of the top layer chip 4 and middle layer chip 5 and non-side inlet opening
Location hole 19 is provided at 18 positions, and the quantity of the location hole 19 is 2 or 4；The top layer chip 4 is 5- high
10mm, middle layer chip 5 are 2-5mm high.
6 material of basal layer chip is in dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate
One kind, 4 material of top layer chip is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate,
5 material of middle layer chip is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate.
A kind of production method of high throughput micro particles circulation sorting and enrichment facility, method includes the following steps:
1) multiple-layer stacked micro-fluidic chip 3 is made:
1. preparing top layer chip 4, middle layer chip 5 and basal layer chip 6；
2. top layer chip 4, middle layer chip 5 are accurately bonded by location hole 19, so that each layer of spiral is nested sinusoidal
About 15 overlapping alignment of runner, opened on the inside of each layer of nested sinusoidal 15 outlet end of runner of spiral from top to bottom later in go out
Fluid hole 16, obtains bonding structure over fluid apertures 17, outside；
3. bonding structure is finally bonded to 6 upper surface of basal layer chip, it is put into baking oven heating later, obtains multiple-layer stacked
Micro-fluidic chip 3；
2) side inlet opening 18 is opened in 3 side of multiple-layer stacked micro-fluidic chip, makes its sinusoidal runner 15 nested with spiral
Liquid feeding end communicates, and is tightly connected with the macropore 21 of flow dispensing connector 2.
3) be sequentially connected each component using connection hose 10~14, constitute the high-throughput micro particles circulation sorting with
Enrichment facility.
1. specific step is as follows for prepare top layer chip 4, middle layer chip 5 and the basal layer chip 6 for the step:
Firstly, in the silicon wafer surface spin coating negative photoresist of cleaned drying and processing, and utilize the photoetching technique of printing film exposure mask
Make the formpiston of top layer chip 4, middle layer chip 5 and basal layer chip 6；Secondly, top layer chip 4, middle layer chip 5 will be prepared
It is poured on the anode membrane prepared with the material of basal layer chip 6, is put into baking oven heating；Finally, demoulding obtain top layer chip 4,
Middle layer chip 5 and basal layer chip 6.
Platymonas helgolandica var cell sorting concentration process is as follows:
Platymonas helgolandica var algae is cultivated, and Platymonas helgolandica var cell is matched by certain concentration with deionized water,
To meet requirement of experiment.Sample containing 100ml Platymonas helgolandica var is put into inner outlet solution receiving flask 8, connects check valve
The scale that connection hose 13 puts in receiving flask is 25ml, imported into flow dispensing connector 2 by peristaltic pump 1 with specific flow
In, flow set range is (500-3500) μ l/min；Sample enters multiple-layer stacked micro-fluidic chip 3 through flow dispensing connector 2；
Platymonas helgolandica var in sample will be by fluid drag force F1Effect and moved along flow direction；Due to howl leaf primary in runner
The parabola shaped velocity profile 23 of stream, microalgae will be by the shear-induced inertia lift F for being directed toward wall surface2；When microalgae leans near wall
When, because the symmetrical tail that rotation rotation generates is generated wall surface induction inertia lift F by Wall effect3；When fluid is by curved
The opposite vortex in two direction of rotation, referred to as Dean stream 24, since Dean flows are generated when runner in vertical main flow direction
24 introducing and generate Dean drag F4；The net inertia lift effect that microalgae is not only generated by inertia migration effect in curved runner
And migrate, also by Dean drag F caused by Secondary Flow4It influences；Therefore, microalgae can be approximately considered in Dean drag F4With it is used
Property lift coupled in common effect under migrate to close to helical flow path inner wall side；Most microalgaes flow into inner outlet solution
Receiving flask 8 then passes through check valve 9 and is again introduced into next circulation that peristaltic pump 1 carries out sorting concentration.When inner outlet solution is received
When collecting 8 surplus 25ml solution of bottle, it is to complete sorting concentration that peristaltic pump 1, which can not continue sample introduction,.
The sorting of high pass Platymonas helgolandica var is recycled in the present embodiment and enrichment facility structure conjunction manufacturing process is simple, and process can
Control, using novel flow channel structure compared with traditional more stable safety of runner, can sub-elect 5ml sample within 3min, and
With intimate 100% sorting index.The plenty of time is saved for sorting enriching service, this technology can be walked by laboratory testing
It has great significance to instant detection.
The above is only the preferred embodiments of the invention, it should be pointed out that: those skilled in the art are come
It says, without departing from the principle of the present invention, can also be adjusted to each facility locations, these adjustment also should be regarded as this hair
Bright protection scope.
Claims (8)
1. a kind of high throughput micro particles circulation sorting and enrichment facility, it is characterised in that: the device includes peristaltic pump (1), stream
Measure dispensing connector (2), multiple-layer stacked micro-fluidic chip (3), outer vent solution receiving flask (7), inner outlet solution receiving flask (8),
Check valve (9) and connection hose (10~14), in which:
The inner outlet solution receiving flask (8) is also initial sample liquid when micro particles circulation sorting starts with concentration process simultaneously
Placement bottle；
By connection hose (13) connection inner outlet solution receiving flask (8), the other end is soft by connecting for described check valve (9) one end
The import of (14) connection peristaltic pump (1) is managed, liquid can only flow to peristaltic pump (1) by inner outlet solution receiving flask (8)；
One end of the flow dispensing connector (2) has macropore (21), and the other end has aperture (20), and the aperture (20) is logical
It crosses connection hose (10) to be connected with the outlet of peristaltic pump (1), the side of the macropore (21) and multiple-layer stacked micro-fluidic chip (3)
Inlet opening (18) is connected directly；
The multiple-layer stacked micro-fluidic chip (3) is from top to bottom by top layer chip (4), middle layer chip (5) and basal layer chip
(6) bonding forms, wherein at least one layer of middle layer chip (5), the contact of the top layer chip (4) and middle layer chip (5)
The contact surface of facial position, the contact surface position of adjacent middle layer chip (5) and middle layer chip (5) and basal layer chip (6)
The nested sinusoidal runner (15) of spiral and side inlet opening (18) are contained in the corresponding position at position, wherein the nested sinusoidal runner of spiral
(15) liquid feeding end is located at the outmost turns of helical structure, and communicates with side inlet opening (18), the nested sinusoidal runner (15) of spiral
Outlet end is located at the innermost circle of helical structure, is provided with interior fluid hole in the interior direction of nested sinusoidal runner (15) outlet end of spiral
(17), the outlet end of the nested sinusoidal runner (15) of the interior fluid hole (17) and spiral communicates, and by connection hose (12) with
Inner outlet solution receiving flask (8) is connected, and is provided with outgoing fluid apertures in the lateral direction of nested sinusoidal runner (15) outlet end of spiral
(16), the outlet end of the nested sinusoidal runner (15) of the outgoing fluid apertures (16) and spiral communicates, and by connection hose (11) with
Outer vent solution receiving flask (7) is connected.
2. a kind of high-throughput micro particles circulation sorting as described in claim 1 and enrichment facility, it is characterised in that: described
Peristaltic pump (1), flow dispensing connector (2), multiple-layer stacked micro-fluidic chip (3), outer vent solution receiving flask (7), inner outlet are molten
Connection between liquid receiving flask (8), check valve (9) and connection hose (10~14) is to be tightly connected.
3. a kind of high-throughput micro particles circulation sorting as described in claim 1 and enrichment facility, it is characterised in that: described
The lower surface of middle layer chip (5) has nested sinusoidal runner (15) structure of concave spiral and concave side inlet opening (18) knot
Structure, upper surface are burnishing surface；The structure of the top layer chip (4) is identical with the structure of middle layer chip (5), and its thickness is greater than
The thickness of middle layer chip (5)；Two surfaces up and down of the basal layer chip (6) are burnishing surface.
4. a kind of high-throughput micro particles circulation sorting as claimed in claim 3 and enrichment facility, it is characterised in that: described
The shape of the nested sinusoidal runner (15) of spiral is to be superimposed sine curve on helical curve, by the nested sinusoidal stream of the concave spiral
Road (15) structure and the burnishing surface of adjacent core lamella are bonded, and the section of the nested sinusoidal runner (15) of the spiral is rectangle,
Width of flow path is 500-700 μm, and runner height is 50 μm -200 μm.
5. a kind of high-throughput micro particles circulation sorting as described in claim 1 and enrichment facility, it is characterised in that: described
Corresponding non-helical nested sine runner (15) and non-side inlet opening (18) position of top layer chip (4) and middle layer chip (5)
Place is provided with location hole (19)；Described top layer chip (4) the high 5-10mm, middle layer chip (5) high 2-5mm.
6. a kind of high-throughput micro particles circulation sorting as described in claim 1 and enrichment facility, it is characterised in that: the base
Bottom chip (6) material is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate, top layer
Chip (4) material is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate, middle layer core
Piece (5) material is one of dimethyl silicone polymer, glass, polycarbonate or polymethyl methacrylate.
7. a kind of production method of high-throughput micro particles circulation sorting and enrichment facility as described in claim 1, feature
It is: method includes the following steps:
1) multiple-layer stacked micro-fluidic chip (3) are made:
1. preparing top layer chip (4), middle layer chip (5) and basal layer chip (6)；
2. top layer chip (4), middle layer chip (5) are accurately bonded by location hole (19), so that each layer of spiral nesting is just
String runner (15) overlapping alignment up and down, later from top to bottom in the inside of each layer nested sinusoidal runner (15) outlet end of spiral
Fluid hole (16) over interior fluid hole (17), outside is opened, bonding structure is obtained；
3. bonding structure is finally bonded to basal layer chip (6) upper surface, it is put into baking oven heating later, it is micro- to obtain multiple-layer stacked
Fluidic chip (3)；
2) side inlet opening (18) are opened in multiple-layer stacked micro-fluidic chip (3) side, makes its sinusoidal runner (15) nested with spiral
Liquid feeding end communicate, and with the macropore (21) of flow dispensing connector (2) be tightly connected；
3) be sequentially connected each component using connection hose (10-14), constitute the high-throughput micro particles circulation sorting with it is dense
Compression apparatus.
8. the production method of a kind of high-throughput micro particles circulation sorting and enrichment facility as claimed in claim 7, feature
Be: 1. the specific steps for preparing top layer chip (4), middle layer chip (5) and basal layer chip (6) are such as the step
Under: firstly, in the silicon wafer surface spin coating negative photoresist of cleaned drying and processing, and utilize the photoetching skill of printing film exposure mask
Art makes the formpiston of top layer chip (4), middle layer chip (5) and basal layer chip (6)；Secondly, will prepare top layer chip (4),
The material of middle layer chip (5) and basal layer chip (6) is poured on the anode membrane prepared, is put into baking oven heating；Finally, demoulding
Obtain top layer chip (4), middle layer chip (5) and basal layer chip (6).
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| CN108160126B (en) * | 2017-11-30 | 2020-05-19 | 东南大学 | Micro-fluidic chip for high-throughput enrichment of micro-particles |
| CN109622078B (en) * | 2018-12-11 | 2020-09-22 | 西安交通大学 | Micro-fluidic chip for single-position enrichment of particles in non-Newtonian fluid |
| CN109975265B (en) * | 2019-04-22 | 2020-06-16 | 中国矿业大学 | Three-dimensional contraction and expansion microfluidic device and method for multidirectional induced Dean flow |
| CN112903411B (en) * | 2021-01-25 | 2023-02-03 | 东南大学 | Multi-mode biological particle concentrator |
| CN115637257B (en) * | 2022-12-07 | 2023-04-14 | 翔鹏佑康(北京)科技有限公司 | Circulating tumor cell screening method based on inertial focusing microfluidics |
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