CN108823065A - Microparticle sorting unit based on intermittent inclination surface acoustic wave - Google Patents
Microparticle sorting unit based on intermittent inclination surface acoustic wave Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
Abstract
The invention discloses a kind of microparticle sorting units based on intermittent inclination surface acoustic wave, including substrate, the cover plate being set on the substrate, the fluid channel for being set to the cover plate bottom and the surface acoustic wave generator for being used to generate inclination surface acoustic wave being set on the substrate.The present invention provides a kind of efficient separation equipment of micro-/ nano particle, the soft lithography manufacture for the standard of can be used, this low cost, high efficiency and portable screening installation can be used for many applications, such as blood/cell/particle separation, cell/particulate media exchange and cell/particle rich.Overall structure sterile sealing of the present invention, to there are the samples of biohazard can also be applicable in；It is designed as micro-fluidic chip plug and play, without cleaning, avoids the cross contamination of each sample room, discardable sorting chip after disposable；System of the invention is not damaged to cell, and provides convenient interface to sample preparation after downstream cellular sorting or the integration of analysis module.
Description
Technical field
It is the present invention relates to micro-fluidic chip, biologic grain detection and manipulation technical field, in particular to a kind of based on interval
The microparticle sorting unit of formula inclination surface acoustic wave.
Background technique
The efficiently separating of suspended particulate and cell studies many basic biomedicals most important, such as cancer cell detection
And drug screening.The cell isolation method most popular in laboratory is density gradient centrifugation at present, according to the size of cell
It is separated with density.The shortcomings that such method, is that the cell that physical property is similar but biological nature is different cannot be distinguished.Separately
It is Fluorescence-activated cell sorting (FACS) that one, which is widely used in industry and the method for clinical field,.Specifically, utilization is glimmering
The monoclonal antibody of signal identifies cell Specific Antigen, will by vibration jet then when by sheath fluid package flow chamber
Liquid stream is converted into the drop comprising individual cells, and drop issues scattered signal and specific fluorescent signal when lining up to pass through laser beam,
Sorting target is set by streaming figure, object droplet is filled with charge in formal sampling, deflects under the action of high voltage electric field
It falls into collection vessel, realizes the separation of cell.In the past few years, chip lab has pushed the hair of cell separation technology
Exhibition, principle include magnetic field, fluid dynamic, luminous power, electrophoresis/dielectrophoresis and acoustic method.
After magnetic field method is incubated for using magnetic bead and cell, under the action of externally-applied magnetic field, by magnetic cell with
Not magnetic cell separates, to reach sorting purpose.Marked by magnetic bead step usually will increase cost and processing time, and may
There is negative effect to cell, the cell bad for antigentic specificity is difficult to realize multiple labelling.Hydrodynamic method is usual
It is related to higher flow velocity or asymmetric barrier.But the obstacle in runner may apply higher mechanical stress and be led to cell
Cause flux decline.Optical tweezer provides a kind of separation method that cell is distinguished according to optical property.However, there are two this methods
Disadvantage：Induced with laser heating, the Multiphoton Absorbtion of the formation of singlet oxygen, biomaterial may result in cell and other biological
The physiological damage of body；This method depends on the optical device of complex and expensive, it is difficult to maintenance and miniaturization.Based on electrophoresis/dielectrophoresis
Method depend on particle polarizability and dielectric conductance rate, due to caused by electric current heating or direct electric field action, may
Have an adverse effect to cell.
Compared with other method for separating, the particle method of operating based on acoustics is non-invasive and is suitable for most of
Microparticle.Bulk acoustic wave (BAW) micro-fluidic chip has shown that good separating effect, however this method needs channel material to have
Excellent sound reflection characteristics (such as silicon and glass).Widely used high molecular material, such as PDMS, do not have usually in microfluidic applications
This standby attribute.Micro-fluidic chip based on surface acoustic wave effectively prevents this limitation, occurs based on sound standing surface wave
(SSAW) micro-fluidic device.In order to increase offset of the microparticle in sorting, oblique interdigital sound standing surface wave cell sorting
Chip is suggested (2014/0033808 A1 of US), and particle is in the region SSAW by acoustic radiation force, resistance, gravity and buoyancy.Weight
Power is similar but contrary with buoyancy size, almost balances.The behavior of particle is radiation fields power and fluid resistance in channel
The coefficient result of power.Acoustic radiation force and resistance can be expressed as：
Fd=-6 π η Rp(up-uf) (3)
P0, Vp in formula, λ, k, x, ρ p, ρ f, β p, β f, η, Rp, up, uf be respectively acoustic pressure, particle volume, wavelength, wave number,
With at a distance from node, grain density, fluid density, particles compress coefficient, fluid compressibility, fluid viscosity, particle radius,
Grain speed, fluid velocity.
Fluid resistance is related with fluid velocity and particle speed.In the inclination stage, due to standing wave nodel line direction and fluid side
To inconsistent, when particle is moved along standing wave nodel line, it will receive the active force of fluid and deviate nodel line, until reaching next section
Line is so unevenly distributed within the scope of width of flow path due to flow velocity, and the position that will result in particle is unpredictable, fluid not
Stablize or is become apparent when having pulsation.In addition, being unfavorable for sorting the control of offset in high flow rate.It is tiltedly interdigital using two-stage
Or tiltedly interdigital timesharing driving, still remain the sound field down periods uncertain problem of particle motion state, especially in high speed or
Person's fluid, which has to embody when pulsation, to be become apparent.
The oblique interdigital increase for being conducive to offset, but since offset is gradually increased with axial displacement, shift time also has
Increased.Shift time is reduced in order to increase offset, interdigital angle and flow rate of liquid are required to increase.But it needs to examine simultaneously
Fluid resistance is considered to the interference effect of motion profile, and interdigital gradient should not be too large, while driving voltage is also required to increase accordingly.
Summary of the invention
In view of the above-mentioned deficiencies in the prior art, the technical problem to be solved by the present invention is that providing a kind of based on interval
The microparticle sorting unit of formula inclination surface acoustic wave.
The tiltedly interdigital zone of action is split by the present invention using directly interdigital, and each oblique interdigital zone of action is smaller,
It is short by the flow interference time when particle is moved along oblique line, it can remain and be moved along nodel line, while offset increases again
It can be effectively controlled deviation post.
In order to solve the above technical problems, the technical solution adopted by the present invention is that：One kind being based on intermittent inclination surface acoustic wave
Microparticle sorting unit, including substrate, the cover plate being set on the substrate, the fluid channel and setting for being set to cover plate bottom
In the surface acoustic wave generator for being used to generate inclination surface acoustic wave on the substrate；The surface acoustic wave generator generates two-way
Standing wave, and the formation sound interacting with surface standing wave area in the fluid channel are superposed to after surface acoustic wave；
Wherein, the sample in the fluid channel is in direction flowing that is single-row and being parallel to fluid channel, and passes through the sound table
Face standing wave active region；
Wherein, the nodel line of sound standing surface wave or anti-nodel line are in intermittent stagger mode parallel and at set angle with sample flow
Shape.
Preferably, it is additionally provided with intermediate between the substrate and cover plate, is provided between the substrate and intermediate
Oxidant layer is coupled, the cover plate is bonded with intermediate.
Preferably, interdigital electrode is set on the substrate and constitutes the surface acoustic wave generator, the interdigital electrode is extremely
Few includes a pair for being symmetricly set on the fluid channel two sides；Between a pair interdigital electrode in the fluid channel two sides
Spacing be the electrode at generate surface acoustic wave half-wavelength integral multiple.
Preferably, the interdigital electrode is in the intermittent cross structure parallel and angled with sample flow, to produce
Raw intermittent inclination surface acoustic wave.
Preferably, the interdigital electrode is fixed width or gradual change width structure.
Preferably, the bottom of the cover plate offers plumbing chase and is connected to the plumbing chase and penetrates through the lid
Multiple apertures of piece side.
Preferably, the substrate is piezoelectricity material, and the cover plate is plastics or glass material；The cover plate sealing attaches
Sealing space and the miniflow is formed on the substrate, making to become in the plumbing chase between the cover plate and piezoelectric substrate
Road.
Preferably, the aperture includes 5, and 3 therein form the sample being connected to one end of the fluid channel and enter
Mouth and the two sheath fluid entrances in sample inlet two sides, in addition 2 form 2 be connected to the other end of the fluid channel out
Mouthful.
Preferably, the width of the fluid channel is 10~1000 μm, is highly 10~500 μm, the diameter of sample
No more than 100um.
Preferably, it is additionally provided with laser detection screening area in the fluid channel of sound interacting with surface standing wave area front end, with
Only one particle to be selected passes through in fluid channel in holding sound interacting with surface standing wave area.
The beneficial effects of the invention are as follows：
The present invention provides a kind of efficient separation equipment of micro-/ nano particle, and the soft lithography manufacture of standard can be used,
This low cost, high efficiency and portable screening installation can be used for many applications, if blood/cell/particle separates, cell/
The exchange of grain culture medium and cell/particle rich.
The prefocus of sample uses hydrodynamic technology in the present invention, makes particle in single-row movement, when guaranteeing follow-up separation
Target particles are moved along a certain fixed nodel line；Intermittent tiltedly each of interdigital oblique interdigital zone of action is smaller, in particle along tiltedly
It is short by the flow interference time when line is mobile, it can remain and be moved along nodel line, offset can be effectively controlled again while increase
Deviation post.
Microparticle sorting unit overall structure sterile sealing of the invention, to there are the samples of biohazard can also be applicable in；
It is designed as micro-fluidic chip plug and play, without cleaning, avoids the cross contamination of each sample room, can after disposable
Abandon sorting chip；System of the invention is not damaged to cell, and to sample preparation or analysis module after downstream cellular sorting
Integration provides convenient interface.
Detailed description of the invention
Fig. 1 is the theory structure schematic diagram of the microparticle sorting unit of the invention based on intermittent inclination surface acoustic wave；
Fig. 2 is the structural schematic diagram of the cover plate in a kind of embodiment of the invention；
Fig. 3 is the knot of the microparticle sorting unit based on intermittent inclination surface acoustic wave of another embodiment of the invention
Structure schematic diagram
Fig. 4 is the structural schematic diagram of the interdigital electrode pair in a kind of embodiment of the invention；
Fig. 5 is that the sample in a kind of embodiment of the invention sorts offset effect figure；
Fig. 6 is that the sample in another embodiment of the invention sorts offset effect figure；
Fig. 7 is that the sample in another embodiment of the invention sorts offset effect figure；
Fig. 8 is that the sample in another embodiment of the invention sorts offset effect figure；
Fig. 9 is the grading principle schematic diagram in another embodiment of the invention.
Description of symbols：
11-the first interdigital electrode；12-the second interdigital electrode；20-fluid channels；21-the first sheath fluid entrance；22-samples
Product entrance；23-the second sheath fluid entrance；24-first outlets；25-second outlets；40-cover plates；41-plumbing chases；42—
First aperture；43-the second aperture；44-third apertures；45-the four aperture；46-the five aperture；50-substrates；51-couplings
Mixture layer；52-intermediates
Specific embodiment
The present invention will be further described in detail below with reference to the embodiments, to enable those skilled in the art referring to specification
Text can be implemented accordingly.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein are not precluded one or more
The presence or addition of a other elements or combinations thereof.
As shown in Figure 1, a kind of microparticle sorting unit based on intermittent inclination surface acoustic wave of the present embodiment, including base
Piece, the cover plate 40 being set on substrate, the fluid channel 20 being set between substrate and cover plate 40 and being set on substrate are used for
Generate the surface acoustic wave generator of inclination surface acoustic wave；Surface acoustic wave generator is superposed to standing wave after generating two channel surface acoustic waves,
And the formation sound interacting with surface standing wave area in fluid channel 20；Sample in fluid channel 20 is in side that is single-row and being parallel to fluid channel 20
To flowing, and pass through sound interacting with surface standing wave area；The nodel line of sound standing surface wave or anti-nodel line are in parallel with sample flow and at settings
The intermittent staggeredly shape of angle.
In one embodiment, coating interdigital electrode constitutes surface acoustic wave generator on substrate, and interdigital electrode includes at least
It is symmetricly set on a pair of 20 two sides of fluid channel, the first interdigital electrode 11 and the second interdigital electrode 12.In 20 two sides of fluid channel
A pair of of interdigital electrode between spacing be the electrode at generate surface acoustic wave half-wavelength integral multiple interdigital electrode be in
The parallel and angled intermittent cross structure of sample flow, to generate intermittent inclination surface acoustic wave.Tilt surface acoustic wave
In inclination be defined as between 0 to 90 degree and 90 to the angle between 180 degree, and do not include 0,90,180 degree.
Wherein, interdigital electrode is fixed width or gradual change width structure.The formula that broadens is interdigital can to generate certain frequency range
Interior sound field (frequency range is determined by interdigital width range), suitable for being used when multichannel sorting.
In one embodiment, in order to promote the recycling rate of waterused of substrate, use cost is reduced, can be used as shown in Figure 3
Structure, daubing coupling agent forms coupling oxidant layer 51 on substrate 50, above attached intermediate 52, cover plate 40 is bonded with intermediate 52.
Discardable cover plate 40 and intermediate 52, substrate 50 are reusable after disposable.Intermediate 52 preferably uses glass or silicon
The small material of acoustic attenuation as piece.Ultrasonic coupling agent and similar acoustic conductive media can be used in couplant, and is applied to sound field work
Use region.
Referring to Fig. 4, in one embodiment, interdigital electrode is to the structure for width gradual change formula.
In one embodiment, referring to Fig. 2, the bottom of cover plate 40 offers plumbing chase 41 and is connected to plumbing chase 41
And 5 apertures of 40 side of cover plate are penetrated through, the first aperture 42, the second aperture 43, third aperture 44, the 4th aperture the 45, the 5th are opened
Hole 46.The sealing of cover plate 40 is attached on substrate, makes to become sealing sky in the plumbing chase 41 between cover plate 40 and piezoelectric substrate
Between and form fluid channel 20.Fluid channel 20 is placed between two pairs of surface acoustic wave generators, and passes through sound interacting with surface standing wave area.It opens
Hole includes 5, and 3 therein form the sample inlet 22 being connected to one end of fluid channel 20 and in 22 two sides of sample inlet
Two sheath fluid entrances, the first sheath fluid entrance 21 and the second sheath fluid entrance 23, in addition 2 formation are connected to the other end of fluid channel 20
2 outlets, first outlet 24 and second outlet 25.Sample inlet 22 in center introduces fluid sample, includes what is manipulated
The sheath fluid entrance of particle, two sides introduces sheath liquid stream, and sample focuses under the package of two sides sheath fluid, in the single-row stream for being parallel to runner
It is dynamic.Fluid in fluid channel remains laminar condition.Packet in the width range of the fluid channel 20 in sound interacting with surface standing wave area
Containing at least one standing wave nodel line (node line) or the anti-nodel line of standing wave (antinode line).Standing wave nodel line present it is parallel with particle stream and
Angled intermittent cross structure.After particle to be selected passes sequentially through sound field reflecting area, by the sound of pressure distribution generation
Radiant force is pushed suspended particulate to pressure node or antinode and is deviated, and the active force that different-grain diameter particle is subject to is different, from
And realize sorting.
Wherein, the material of cover plate 40 is polymer, plastics or glass, and the material of piezoelectric substrate is piezoelectric ceramics, piezoelectricity
Monocrystalline or piezo-electricity composite material.
Wherein, fluid channel 20 may include a plurality of, to carry out multiple groups sorting simultaneously.The width of fluid channel 20 is 10~1000
μm, be highly 10~500 μm, the diameter of sample is not more than 100um.It further include droplet and cell in sample
Wherein, the voltage waveform applied in interdigital electrode is sine wave or square wave, if desired according to detection signal to particle
Targeting screening is carried out, pulse envelope signal can be applied after detecting echo signal, such as the sine wave or square wave of square wave envelope.
Wherein, there is great role in the position that sample flow enters the region SSAW for offset and sorting success or failure, is shown in Fig. 5
Preferred embodiment situation, the sample flow after focusing are overlapped with nodel line 30c, and particle will be moved along nodel line when sound field reflecting,
It is less by the lesser little particle offset of acoustic radiation force, two kinds of particles can be separated in subsequent fork level.In another embodiment
In, as Fig. 6 will result in the damage of offset if there is certain deviation in the position that sample enters sound field reflecting area relative to nodel line 32c
It loses, is likely to result in follow-up separation failure.In another embodiment, such as Fig. 7, if sample enters the position phase in sound field reflecting area
It is closely located for two nodel lines 34b, 34c, it will result in the uncertainty of target offset, it is also possible to will cause follow-up separation mistake
Effect.In another embodiment, still cannot after offset such as Fig. 8 if the nodel line 36a selection in sound field reflecting area is improper
Target particles are separated, it is also possible to will cause follow-up separation failure.
Increase in the runner of the region SSAW (sound interacting with surface standing wave area) front end in another embodiment referring to Fig. 9
Laser detection area 50 carries out targeting screening to particle according to detection signal, keeps in the region SSAW 51 that only one is to be selected always
Particle passes through, and if object, then applies pulsed ultrasonic field and selects, sound field is otherwise not added and is allowed to pass freely through, to further increase
Separating effect.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details.
Claims (10)
1. a kind of microparticle sorting unit based on intermittent inclination surface acoustic wave, which is characterized in that including substrate, be set to institute
State the cover plate on substrate, be set to the fluid channel of cover plate bottom and be set on the substrate for generate inclination surface acoustic wave
Surface acoustic wave generator；The surface acoustic wave generator is superposed to standing wave after generating two channel surface acoustic waves, and in the miniflow
Formation sound interacting with surface standing wave area on road；
Wherein, the sample in the fluid channel is in direction flowing that is single-row and being parallel to fluid channel, and is stayed by the sound surface
Wave active region；
Wherein, the nodel line of sound standing surface wave or anti-nodel line are in intermittent staggeredly shape parallel with sample flow and at set angle.
2. the microparticle sorting unit according to claim 1 based on intermittent inclination surface acoustic wave, which is characterized in that institute
State and be additionally provided with intermediate between substrate and cover plate, be provided with coupling oxidant layer between the substrate and intermediate, the cover plate with
Intermediate bonding.
3. the microparticle sorting unit according to claim 1 based on intermittent inclination surface acoustic wave, which is characterized in that institute
It states and the interdigital electrode composition surface acoustic wave generator is set on substrate, the interdigital electrode is described including at least being symmetricly set on
A pair of fluid channel two sides；Spacing between a pair interdigital electrode in the fluid channel two sides is to generate at the electrode
Surface acoustic wave half-wavelength integral multiple.
4. the microparticle sorting unit according to claim 3 based on intermittent inclination surface acoustic wave, which is characterized in that institute
Interdigital electrode is stated in the intermittent cross structure parallel and angled with sample flow, to generate intermittent inclination sound surface
Wave.
5. the microparticle sorting unit according to claim 3 based on intermittent inclination surface acoustic wave, which is characterized in that institute
Stating interdigital electrode is fixed width or gradual change width structure.
6. the microparticle sorting unit according to claim 1 based on intermittent inclination surface acoustic wave, which is characterized in that institute
The bottom for stating cover plate offers plumbing chase and multiple apertures of the cover plate side is connected to and penetrated through with the plumbing chase.
7. the microparticle sorting unit according to claim 6 based on intermittent inclination surface acoustic wave, which is characterized in that institute
Stating substrate is piezoelectricity material, and the cover plate is plastics or glass material；The cover plate sealing is attached on the substrate, makes to be in
Plumbing chase between the cover plate and piezoelectric substrate becomes sealing space and forms the fluid channel.
8. the microparticle sorting unit according to claim 7 based on intermittent inclination surface acoustic wave, which is characterized in that institute
Stating aperture includes 5, and 3 therein form the sample inlet being connected to one end of the fluid channel and in sample inlet two sides
Two sheath fluid entrances, in addition 2 form be connected to the other end of the fluid channel 2 and export.
9. the microparticle sorting unit according to claim 1 based on intermittent inclination surface acoustic wave, which is characterized in that institute
The width for stating fluid channel is 10~1000 μm, is highly 10~500 μm, and the diameter of sample is not more than 100um.
10. the microparticle sorting unit according to claim 1 based on intermittent inclination surface acoustic wave, which is characterized in that
It is additionally provided with laser detection screening area in the fluid channel of sound interacting with surface standing wave area front end, to keep sound interacting with surface standing wave
The particle to be selected of only one in fluid channel in area passes through.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109439513A (en) * | 2018-12-11 | 2019-03-08 | 中国科学院苏州生物医学工程技术研究所 | The micro-fluidic chip screened for rare cell in whole blood |
| CN109482121A (en) * | 2018-12-27 | 2019-03-19 | 苏州纳葛诺斯生物科技有限公司 | Micro-and nano-particles highly effective reaction micro-fluidic chip based on surface acoustic wave |
| CN110918140A (en) * | 2018-09-20 | 2020-03-27 | 北京怡天佳瑞科技有限公司 | Microfluidic chip, device containing same and method for separating particles |
| CN111925934A (en) * | 2020-07-31 | 2020-11-13 | 深圳先进技术研究院 | Biological sample sorting method, surface acoustic wave micro-fluidic chip, system, terminal and storage medium |
| CN112080385A (en) * | 2019-06-13 | 2020-12-15 | 安行生物技术有限公司 | Method and apparatus for separating cells or microvesicles |
| CN112492471A (en) * | 2020-12-04 | 2021-03-12 | 西安交通大学 | Acoustic valve device based on micro-fluidic and surface acoustic wave technology |
| CN112986108A (en) * | 2021-03-21 | 2021-06-18 | 厦门大学 | Flow cell for focusing nano particles based on surface acoustic waves |
| CN114556258A (en) * | 2019-04-24 | 2022-05-27 | 班努生物科技公司 | Location-assisted negative particle rejection (PANR) for sorting and enriching target cells of interest |
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