CN107603850A - Micro fluidic device for cell sorting and preparation method thereof - Google Patents
Micro fluidic device for cell sorting and preparation method thereof Download PDFInfo
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- CN107603850A CN107603850A CN201710961288.2A CN201710961288A CN107603850A CN 107603850 A CN107603850 A CN 107603850A CN 201710961288 A CN201710961288 A CN 201710961288A CN 107603850 A CN107603850 A CN 107603850A
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Abstract
The invention provides a kind of micro fluidic device for cell sorting and preparation method thereof, it is related to cell sorting techniques field, this is used for the micro fluidic device of cell sorting, including lower substrate and the upper matrix that be connected with lower substrate, and lower substrate is provided with the micro-pillar array for sorting cell；Flowed to along cell, the orientation of the column in micro-pillar array and the angle of cell flow direction are 2 ° 10 °；Flow to direction along perpendicular to cell, the spacing between column in micro-pillar array is 8 13 microns, using the micro fluidic device can alleviate prior art cell sorting device is complicated and the high technical problem of equipment cost.
Description
Technical field
The present invention relates to cell sorting techniques field, more particularly, to a kind of micro fluidic device for cell sorting and its
Preparation method.
Background technology
Cell sorting is the necessary means that the homogeneous target cell of property is obtained from complex environment.Cell sorting is cell
The key link of physiology and pathological study, its diagnosis and treatment for major disease are significant.For cell sorting
The research of method and correlation technique always is a study hotspot of medical field and association area.
Existing cell sorting mode is mainly by using different fluorescence labeled cells, with the streaming with sorting function
Cell instrument is sorted.Flow cytometer is to be irradiated with high energy laser under flow at high speed state by the slender of fluorochromes
Born of the same parents or particulate, its caused intensity for scattering light and launching fluorescence is measured to separate cell.But the streaming with sorting function
Cell instrument volume is big, complicated, expensive, and cumbersome to the processing mode of cell, and experimental cost is higher.
The content of the invention
The first object of the present invention is to provide a kind of micro fluidic device for cell sorting, to alleviate prior art
Cell sorting device is complicated and the high technical problem of equipment cost.
The second object of the present invention is to provide a kind of preparation method of the micro fluidic device for cell sorting, this method
With manufacturing cost it is low the advantages of.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of micro fluidic device for cell sorting, including lower substrate and the upper matrix that is connected with the lower substrate, institute
State lower substrate and be provided with the micro-pillar array for being used for sorting cell；Flowed to along cell, the arrangement side of the column in the micro-pillar array
It it is 2 ° -10 ° to the angle flowed to the cell；Direction is flowed to along perpendicular to cell, between the column in the micro-pillar array
Spacing be 8-13 microns.
Further, flowed to along cell, the orientation of the column in the micro-pillar array and the folder of cell flow direction
Angle is 2 ° -9 °, more preferably 2 ° -8 °；Flow to direction along perpendicular to cell, between the column in the micro-pillar array between
Away from for 8-12 microns, more preferably 10-12 microns.
Further, the column in the micro-pillar array includes triangular stud, circular abutment or trapezoid stand column.
Further, the triangular stud is isosceles triangle column；Preferably, the isosceles triangle column is straight
Angle isosceles triangle column or obtuse angle isosceles triangle column.
Further, cell flows to side where base when flowing and in the isosceles triangle column along the cell
Collide.
Further, direction is flowed to along perpendicular to cell, the both sides of the lower substrate, which are provided with, to be used to apply magnetic field to cell
Magnetic control means.
Further, direction is flowed to along perpendicular to cell, the both sides of the lower substrate, which are provided with, to be used to apply electric field to cell
Electrode.
Further, flowed to along cell, one end of the lower substrate is provided with the first inlet and use that are used for injecting cell
In the second inlet of injection culture medium；
The other end of the lower substrate is provided with sample export, and the sample export is no less than two and along perpendicular to cell stream
To direction dispersed placement.
A kind of preparation method of the above-mentioned micro fluidic device for cell sorting, first passes through casting method and lower base is prepared
Body, then the lower substrate is connected with upper matrix to obtain the micro fluidic device for being used for cell sorting.
Further, the casting method includes pouring liquid charging stock in mould, isolated institute after cured shaping
State lower substrate；
Preferably, the liquid charging stock includes the prepolymer of curing agent and PDMS compositions；
Preferably, the curing agent and the PDMS weight ratio are 1：(8-12)；
Preferably, degassing process resolidification shaping is first carried out after pouring；
Preferably, the temperature during curing molding is 80-100 DEG C, time 1.5-2.5h.
Further, the preparation method of the mould comprises the following steps：Photoresist is coated on die matrix surface, is passed through
The cavity structure corresponding with the micro-pillar array in the lower substrate is obtained after exposure, development；
Preferably, the thickness of the photoresist of coating is 8-14 microns.
Compared with the prior art, the present invention has the advantages that：
Provided by the present invention for the micro fluidic device of cell sorting, due to micro-pillar array central post orientation with it is thin
Certain angle be present in born of the same parents flow direction, cause the cell of different sizes pass through the angle that is repelled during micro-pillar array and away from
From difference, in the case where flow velocity is certain, the angle and distance that large-sized cell is repelled can be some larger.Cell
After being repelled recycle column between interval can further increase between big cellule flowing difference (including flowing angle with
With the offset distance of cell incident direction), thus in the present invention by rationally set micro-pillar array central post orientation and
Spacing is arranged, makes cell according to its size fatefully selective flow path, it is various various sizes of so as to dexterously realize
The sorting of cell.
Micro fluidic device provided by the invention has simple in construction relative to existing flow cytometer, and cost is low, does not have to
The advantages of sorting to cell can be achieved in fluorescence processing is carried out to cell.
Brief description of the drawings
, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical scheme of the prior art
The required accompanying drawing used is briefly described in embodiment or description of the prior art, it should be apparent that, in describing below
Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before creative work is not paid
Put, other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 is the structural representation for the micro fluidic device for cell sorting that the embodiment of the present invention 1 provides；
Fig. 2 is the structural representation for the micro fluidic device for cell sorting that the embodiment of the present invention 2 provides；
Fig. 3 is the structural representation for the micro fluidic device for cell sorting that invention embodiment 3 provides；
Fig. 4 is the structural representation for the micro fluidic device for cell sorting that invention embodiment 4 provides.
Icon：10- lower substrates；11- micro-pillar arrays；12- columns；The inlets of 13- first；The inlets of 14- second；15- samples
Product export；20- magnetic control means；30- electrodes.
Embodiment
Technical scheme is clearly and completely described below in conjunction with accompanying drawing, it is clear that described implementation
Example is part of the embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, ordinary skill
The every other embodiment that personnel are obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
In the description of the invention, it is necessary to explanation, term " " center ", " on ", " under ", "left", "right", " vertical ",
The orientation or position relationship of the instruction such as " level ", " interior ", " outer " be based on orientation shown in the drawings or position relationship, merely to
Be easy to the description present invention and simplify description, rather than instruction or imply signified device or element must have specific orientation,
With specific azimuth configuration and operation, therefore it is not considered as limiting the invention.In addition, term " first ", " second ",
" the 3rd " is only used for describing purpose, and it is not intended that instruction or hint relative importance.
In the description of the invention, it is necessary to illustrate, unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " should be interpreted broadly, for example, it may be being fixedly connected or being detachably connected, or be integrally connected；Can
To be mechanical connection or electrical connection；Can be joined directly together, can also be indirectly connected by intermediary, Ke Yishi
The connection of two element internals.For the ordinary skill in the art, above-mentioned term can be understood as the case may be
Concrete meaning in the present invention.
One aspect of the present invention provides a kind of micro fluidic device for cell sorting, including lower substrate and with it is described
The upper matrix of lower substrate connection, the lower substrate are provided with the micro-pillar array for being used for sorting cell；Flowed to along cell, the microtrabeculae
The orientation of column in array and the angle of cell flow direction are 2 ° -10 °；Edge flows to direction perpendicular to cell, described
The spacing between column in micro-pillar array is 8-13 microns.
Provided by the present invention for the micro fluidic device of cell sorting, due to micro-pillar array central post orientation with it is thin
Certain angle be present in born of the same parents flow direction, cause the cell of different sizes pass through the angle that is repelled during micro-pillar array and away from
From difference, in the case where flow velocity is certain, the angle and distance that large-sized cell is repelled can be some larger.Cell
After being repelled recycle column between interval can further increase between big cellule flowing difference (including flowing angle with
With the offset distance of cell incident direction), thus in the present invention by rationally set micro-pillar array central post orientation and
Spacing is arranged, makes cell according to its size fatefully selective flow path, it is various various sizes of so as to dexterously realize
The sorting of cell.
In addition, micro fluidic device provided by the invention has simple in construction relative to existing flow cytometer, cost is low,
Without carrying out the advantages of sorting to cell can be achieved in fluorescence processing to cell.
The middle part of lower substrate is the sorting region being made up of micro-pillar array, and wherein micro-pillar array presses transverse and longitudinal two by column
Individual direction is arranged to make up, and longitudinally represents the flow direction of cell, is laterally the direction perpendicular with the flow direction of cell, thirty years of age
The short transverse of post represents the depth dimension in sorting region.
Micro-pillar array in the present invention, can be one group can also be it is multigroup, when for it is multigroup when can further improve point
The precision of choosing.
The angular range of the orientation of micro-pillar array central post and cell flow direction is 2 ° -10 °, when needs sort not
With size cell when, the angle can be arranged to different sizes.In addition, when micro-pillar array is multigroup, can also incite somebody to action
The micro-pillar array of difference group is arranged to different angles, can also set different angles in same group of micro-pillar array, so may be used
Further to increase separating effect.
Equally, the setting scope of the spacing between micro-pillar array central post is 8-13 microns, when needing to sort different sizes
Cell when, the spacing can be arranged to different sizes.In addition, when micro-pillar array is multigroup, can also be by different groups
Micro-pillar array be arranged to different spacing, different spacing can also be set in same group of micro-pillar array, can so enter one
Step increase separating effect.
Lower substrate is preferably made using PDMS material, and upper matrix preferably uses sheet glass, and both preferably use key
The mode of conjunction combines the micro fluidic device formed for cell sorting.
In the present invention, the orientation of column and the angle of cell flow direction are typical but non-limiting for example,：2°、
3 °, 4 °, 5 °, 6 °, 7 °, 8 °, 9 ° or 10 °；The spacing between column in micro-pillar array is typical but non-limiting for example,：8
Micron, 9 microns, 10 microns, 11 microns, 12 microns or 13 microns.
As the preferred embodiment of the present invention, flowed to along cell, the orientation of the column in the micro-pillar array with
The angle of the cell flow direction is 2 ° -9 °, more preferably 2 ° -8 °；Direction, the micro-pillar array are flowed to along perpendicular to cell
In column between spacing be 8-12 microns, more preferably 10-12 microns.By further optimal design-aside angle and
Away from sharpness of separation can be improved.
As the preferred embodiment of the present invention, the column in the micro-pillar array includes triangular stud, circular abutment
Or trapezoid stand column.Column in micro-pillar array can be the column of various shapes structure, in addition to above-mentioned several shapes, can also be
Other shapes.
As further preferred embodiment of the present invention, the triangular stud is isosceles triangle column；Further
Preferably, the isosceles triangle column is right angled isosceles triangle column or obtuse angle isosceles triangle column.Using isosceles three
Corniform upright column, it can be produced on each side when cell flows to column and collide concurrent hair tonic and penetrate.Using right angle isosceles
Secondary reflection of the cell between column can be reduced when triangle or obtuse angle isosceles triangle, it is reflected by primary collision
After can smoothly between two columns by, meanwhile, the preparation of mould is also convenient for using right angled isosceles triangle.
As the preferred embodiment of the present invention, cell is flowed to along the cell when flowing and the isosceles triangle column
In base where sideways collisions.Now side where the base in isosceles triangle column flows to inconsistent set with cell
Put, cell is flowed to after column in the sideways collisions where base and reflect away.
When using isosceles triangle column, the side where base flows to angle at 45 ° with cell, makes the transmitting of cell more
It is regular.
As the preferred embodiment of the present invention, direction is flowed to along perpendicular to cell, the both sides of the lower substrate, which are provided with, to be used
In the magnetic control means for applying magnetic field to cell.In a preferred embodiment of the present invention, the magnetic control means includes being arranged at lower substrate two
The bar magnet of side, bar magnet, which is powered, can produce magnetic field, and caused magnetic field can control cell to deviate cell flow direction.Now different chis
The magneticaction that very little size or different types of cell are subject to is different, and deviation effect caused by the big cell of stress is more obvious,
Therefore, it can be reached by increasing magnetic field and further divide cellifugal effect.
As the preferred embodiment of the present invention, direction is flowed to along perpendicular to cell, the both sides of the lower substrate, which are provided with, to be used
In the electrode for applying electric field to cell.When containing electrically charged cell in the cell for needing to separate by the way that cell is powered and can divided
Separate out the electrically charged cell in cell.
As the preferred embodiment of the present invention, flowed to along cell, one end of the lower substrate, which is provided with, to be used to inject cell
The first inlet and the second inlet for injecting culture medium；The other end of the lower substrate is provided with sample export, described
Sample export flows to direction dispersed placement no less than two and along perpendicular to cell.
Cell sample entrance is used to connect cell groove, and culture medium inlet is used to connect culture foundation trench groove tank, sample export
For connecting cell collecting tank, because the cell of different sizes or type can be obtained after cell separation, therefore sample export can
It is multiple to set, and sample export flows to the transversely arranged direction in direction, i.e. column in micro-pillar array along edge perpendicular to cell
Dispersed placement.
Another aspect of the present invention provides a kind of preparation method of the above-mentioned micro fluidic device for cell sorting, first
Lower substrate is prepared by casting method, then the lower substrate is bonded with upper matrix to obtain the miniflow for being used for cell sorting
Control device.
Lower substrate in the present invention is prepared by casting method, and its technical process is simple, and is easy to implement, and obtains lower base
It is connected again with upper matrix after body and can obtain the above-mentioned micro fluidic device for cell sorting.Lower substrate and upper matrix in the present invention
Connection preferably using bonding connection.
As the preferred embodiment of the present invention, the casting method includes pouring liquid charging stock in mould, cured
The isolated lower substrate after shaping；Alternatively, the liquid charging stock includes curing agent and dimethyl silicone polymer
(Polydimethylsiloxane, referred to as：PDMS) the prepolymer of composition；Alternatively, the weight of the curing agent and the PDMS
Amount is than being 1：(8-12).In a preferred embodiment of the invention, lower substrate is made using PDMS material, and upper matrix is flat
Glass sheet.
In above-mentioned preferred embodiment, the curing agent and the PDMS weight are than typical but non-limiting example
Such as it is 1:8、1:9、1:10、1:11 or 1:12；Wherein described curing agent is preferably dealcoholized type curing agent or depickling type curing agent.
As the preferred embodiment of the present invention, degassing process resolidification shaping is first carried out after pouring；It is further preferred that
Temperature during curing molding is 80-100 DEG C, time 1.5-2.5h.First carrying out degassing process can exclude in PDMS
Bubble, ensure bubble-free remaining influence in lower substrate, have a negative impact with separation of the Anti-bubble to cell.
In above-mentioned preferred embodiment, the temperature during curing molding is typical but non-limiting for example,：80
DEG C, 85 DEG C, 90 DEG C, 95 DEG C or 100 DEG C；Time during curing molding is typical but non-limiting for example,：1.5h、
1.7h, 2.0h, 2.2h or 2.5h.
As the preferred embodiment of the present invention, the preparation method of the mould comprises the following steps：In die matrix table
Face coats photoresist, by obtaining the cavity structure corresponding with lower substrate structure after exposing, developing；Alternatively, the light of coating
The thickness of photoresist is 8-14 microns.In the preferred embodiment of the present invention, mould is that photoresist is coated on silicon chip, passes through exposure
Obtained after light, development, this method operating procedure is simple, is easy to implement.
In above-mentioned preferred embodiment, the thickness of the photoresist of coating is typical but non-limiting to be, for example,：8 is micro-
Rice, 10 microns, 11 microns, 12 microns, 13 microns or 14 microns.
As the preferred embodiment of the present invention, lower substrate using puncher carries out punching system after completing to lower substrate
For cell sample entrance, culture medium inlet and sample export is gone out, finally it is bonded with upper matrix, obtains the micro fluidic device.
Below in conjunction with embodiment, comparative example and accompanying drawing, the present invention will be further described in detail.
Embodiment 1
As shown in figure 1, the present embodiment is a kind of micro fluidic device for cell sorting, including lower substrate 10 and with lower base
The upper matrix that body 10 connects, upper matrix are slide, and lower substrate 10 is provided with the micro-pillar array 11 for being used for sorting cell, along cell
Flow to, the orientation of the column 12 in micro-pillar array 11 and the angle of cell flow direction are 2.8 °；Along perpendicular to cell flow direction side
To the spacing between column 12 in micro-pillar array 11 is 10 microns.Micro-pillar array 11 in the present embodiment is one group, microtrabeculae battle array
Column 12 in row 11 is circular abutment.Flowed to along cell, one end of lower substrate 10 is provided with cell sample entrance 13 and culture medium
Inlet 14, cell sample entrance 13 are used to connect cell groove, and culture medium inlet 14 is used to connect culture medium groove tank；Lower substrate
10 other end is provided with sample export 15, and sample export 15 is used to connect 3 cell collecting tanks, and sample export 15 is no less than two
And flow to direction dispersed placement along perpendicular to cell.
Wherein, the dotted line line between Fig. 1 central posts 12 shows the orientation of the central post 12 of micro-pillar array 11.
Embodiment 2
The present embodiment is a kind of micro fluidic device for cell sorting, and the difference with embodiment 1 is, along cell stream
To the orientation of the column in micro-pillar array and the angle of cell flow direction are 9 °；Direction, microtrabeculae battle array are flowed to along perpendicular to cell
The spacing between column in row is 12 microns.
Embodiment 3
The present embodiment is a kind of micro fluidic device for cell sorting, and the difference with embodiment 1 is, along cell stream
To the orientation of the column in micro-pillar array and the angle of cell flow direction are 10 °；Direction, microtrabeculae are flowed to along perpendicular to cell
The spacing between column in array is 13 microns.
Embodiment 4
As shown in Fig. 2 the present embodiment is a kind of micro fluidic device for cell sorting, including lower substrate 10 and with lower base
The upper matrix that body 10 connects, lower substrate 10 are provided with the micro-pillar array 11 for being used for sorting cell, flowed to along cell, micro-pillar array 11
In the angle of orientation and cell flow direction of column 12 be 5.6 °；Direction is flowed to along perpendicular to cell, in micro-pillar array 11
Column 12 between spacing be 10 microns.Micro-pillar array 11 in the present embodiment is one group, the column 12 in micro-pillar array 11
For right angled isosceles triangle column.Flowed to along cell, one end of lower substrate 10 is provided with cell sample entrance 13 and culture medium injects
Mouth 14, cell sample entrance 13 are used to connect cell groove, and culture medium inlet 14 is used to connect culture foundation trench groove tank；Lower substrate 10
The other end be provided with sample export 15, sample export 15 is used to connecting 3 cell collecting tanks, sample export 15 no less than two and
Direction dispersed placement is flowed to along perpendicular to cell.Wherein, the dotted line line between Fig. 2 central posts 12 is shown in micro-pillar array 11
The orientation of column 12.
Embodiment 5
As shown in figure 3, the present embodiment is a kind of micro fluidic device for cell sorting, as different from Example 2, this
Micro-pillar array 11 in embodiment is three groups, the orientation of the column 12 in first group of micro-pillar array 11 and the folder of cell flow direction
Angle is that the angle that the orientation of the column 12 in 2.8 °, second group and the 3rd group micro-pillar arrays 11 flows to cell is
5.6°；Direction is flowed to along perpendicular to cell, and the spacing between column 12 in first group of micro-pillar array 11 is 10 microns, second group
Spacing between the column 12 in the 3rd group of micro-pillar array 11 is 12 microns.Wherein, the dotted line between Fig. 3 central posts 12 connects
Line shows the orientation of the central post 12 of micro-pillar array 11.
Embodiment 6
As shown in figure 4, the present embodiment is a kind of micro fluidic device for cell sorting, as different from Example 3, this
Direction is flowed to along perpendicular to cell in embodiment, the both sides of lower substrate 10 are provided with the magnetic control means 20 for being used for applying magnetic field to cell
With the electrode 30 for applying electric field to cell.Wherein, the dotted line line between Fig. 4 central posts 12 is shown in micro-pillar array 11
The orientation of column 12.
Comparative example 1
This comparative example is a kind of micro fluidic device for cell sorting, and the difference with embodiment 1 is, along cell stream
To the orientation of the column in micro-pillar array and the angle of cell flow direction are 1 °；Direction, microtrabeculae battle array are flowed to along perpendicular to cell
The spacing between column in row is 3 microns.
Comparative example 2
This comparative example is a kind of micro fluidic device for cell sorting, and the difference with comparative example 1 is, along cell stream
To the orientation of the column in micro-pillar array and the angle of cell flow direction are 1 °；Direction, microtrabeculae battle array are flowed to along perpendicular to cell
The spacing between column in row is 10 microns.
Comparative example 3
This comparative example is a kind of micro fluidic device for cell sorting, and the difference with comparative example 1 is, along cell stream
To the orientation of the column in micro-pillar array and the angle of cell flow direction are 2.8 °；Direction, microtrabeculae are flowed to along perpendicular to cell
The spacing between column in array is 3 microns.
Comparative example 4
This comparative example is a kind of micro fluidic device for cell sorting, and the difference with embodiment 4 is, along cell stream
To the orientation of the column in micro-pillar array and the angle of cell flow direction are 12 °；Direction, microtrabeculae are flowed to along perpendicular to cell
The spacing between column in array is 15 microns.
Comparative example 5
This comparative example is a kind of micro fluidic device for cell sorting, and the difference with embodiment 5 is, along cell stream
To the orientation of the column in first group of micro-pillar array and the angle of cell flow direction are 1.5 °, second group and the 3rd group microtrabeculaes
The orientation of column in array and the angle of cell flow direction are 12 °；Direction, first group of microtrabeculae are flowed to along perpendicular to cell
Spacing of the spacing between column between the column in 10 microns, second group and the 3rd group micro-pillar arrays in array is 12
Micron.
Comparative example 6
This comparative example is a kind of micro fluidic device for cell sorting, and the difference with embodiment 6 is, first group of microtrabeculae
The angle of the orientation of column in array and cell flow direction is the row of the column in 1 °, second group and the 3rd group micro-pillar array
Column direction and the angle of cell flow direction are 1.2 °；Flow to direction along perpendicular to cell, the column in first group of micro-pillar array it
Between spacing of the spacing between the column in 4 microns, second group and the 3rd group micro-pillar arrays be 5 microns.
Confirmatory experiment：Same cell mass is entered with the micro fluidic device provided in embodiment 1-6 and comparative example 1-6 respectively
Row sorting, specific test operation process are as follows：Bacterium to be separated is grown into logarithmic phase afterwards twice with Shaking culture switching,
During OD600=0.2, come in and gone out with after first syringe absorption bacteria samples from cell sample entrance, while injected with second
Device injects after drawing culture medium from culture medium inlet, stream when bacterium and culture medium being injected and injected simultaneously by peristaltic pump
Speed is identical；Bacterium flows through enters the cell collecting tank being connected with sample export after micro fluidic device through sample export, concentrates on and receives
Micro- sem observation tracking is carried out in collection groove, by the image processing and analyzing means such as matlab, the locomitivity difference of cell can be obtained
Related data.
Experiment proves that 2-4um bacterium can be separated into average length well and be by the micro fluidic device in example 1-6
2 μm, 3 μm and 4 μm three parts, have added the effect of electric field to become apparent, the length of bacteria discrimination of separate collection can be of about 0.9
μm.When carrying out size sorting to Escherichia coli using the micro fluidic device in comparative example 1-6, because inclination angle is too small or spacing mistake
Wide reason, it is impossible to well separate bacterium.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to foregoing embodiments for pipe, it will be understood by those within the art that：Its according to
The technical scheme described in foregoing embodiments can so be modified, either which part or all technical characteristic are entered
Row equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention technology
The scope of scheme.
Claims (10)
1. a kind of micro fluidic device for cell sorting, it is characterised in that be connected including lower substrate and with the lower substrate
Upper matrix, the lower substrate are provided with the micro-pillar array for being used for sorting cell；Flowed to along cell, the column in the micro-pillar array
The angle of orientation and the cell flow direction be 2 ° -10 °；Direction is flowed to along perpendicular to cell, in the micro-pillar array
Spacing between column is 8-13 microns.
2. the micro fluidic device according to claim 1 for cell sorting, it is characterised in that flowed to along cell, it is described
The orientation of column in micro-pillar array and the angle of cell flow direction are 2 ° -9 °, more preferably 2 ° -8 °；Hang down on edge
Directly direction is flowed in cell, the spacing between column in the micro-pillar array is 8-12 microns, and more preferably 10-12 is micro-
Rice.
3. the micro fluidic device according to claim 1 for cell sorting, it is characterised in that in the micro-pillar array
Column includes triangular stud, circular abutment or trapezoid stand column；
Preferably, the triangular stud is isosceles triangle column；
Preferably, the isosceles triangle column is right angled isosceles triangle column or obtuse angle isosceles triangle column.
4. the micro fluidic device according to claim 3 for cell sorting, it is characterised in that cell is along the cell stream
To sideways collisions where base during flowing and in the isosceles triangle column.
5. according to the micro fluidic device for cell sorting described in claim any one of 1-4, it is characterised in that along perpendicular to
Cell flows to direction, and the both sides of the lower substrate are provided with the magnetic control means for being used for applying magnetic field to cell.
6. according to the micro fluidic device for cell sorting described in claim any one of 1-4, it is characterised in that along perpendicular to
Cell flows to direction, and the both sides of the lower substrate are provided with the electrode for being used for applying electric field to cell.
7. according to the micro fluidic device for cell sorting described in claim any one of 1-4, it is characterised in that along cell stream
To one end of the lower substrate, which is provided with, is used for the first inlet for injecting cell and the second inlet for injecting culture medium；
The other end of the lower substrate is provided with sample export, and the sample export is no less than two and along perpendicular to cell flow direction side
To dispersed placement.
8. a kind of preparation method of the micro fluidic device for cell sorting described in any one of claim 1-7, its feature exist
In first passing through casting method and be prepared lower substrate, then the lower substrate is connected with upper matrix obtain and described is used for cell sorting
Micro fluidic device.
9. preparation method according to claim 8, it is characterised in that the casting method includes pouring liquid charging stock in mould
In tool, the isolated lower substrate after cured shaping；
Preferably, the liquid charging stock includes the prepolymer of curing agent and PDMS compositions；
Preferably, the curing agent and the PDMS weight ratio are 1：(8-12)；
Preferably, degassing process resolidification shaping is first carried out after pouring；
Preferably, the temperature during curing molding is 80-100 DEG C, time 1.5-2.5h.
10. preparation method according to claim 9, it is characterised in that the preparation method of the mould comprises the following steps：
Photoresist is coated on die matrix surface, it is corresponding with the micro-pillar array in the lower substrate by being obtained after exposing, developing
Cavity structure；
Preferably, the thickness of the photoresist of coating is 8-14 microns.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710961288.2A CN107603850A (en) | 2017-10-13 | 2017-10-13 | Micro fluidic device for cell sorting and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710961288.2A CN107603850A (en) | 2017-10-13 | 2017-10-13 | Micro fluidic device for cell sorting and preparation method thereof |
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| Publication Number | Publication Date |
|---|---|
| CN107603850A true CN107603850A (en) | 2018-01-19 |
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| CN109097245A (en) * | 2018-07-25 | 2018-12-28 | 大连理工大学 | A kind of new microfluidic array arrested for cell high-efficient |
| CN110420672A (en) * | 2019-07-16 | 2019-11-08 | 北京化工大学 | It a kind of micro-fluidic chip and its cleans in particle and changes the application in liquid |
| CN112029662A (en) * | 2020-11-06 | 2020-12-04 | 深圳市赛特罗生物医疗技术有限公司 | Cell sorting magnetic grid structure, manufacturing method and magnetic grid tube |
| CN112094839A (en) * | 2020-11-06 | 2020-12-18 | 深圳市赛特罗生物医疗技术有限公司 | Automatic cell magnetic sorting method and device |
| CN112553043A (en) * | 2020-12-09 | 2021-03-26 | 深圳先进技术研究院 | Microfluidic chip for separating and purifying fetal nucleated red blood cells |
| CN112574851A (en) * | 2019-09-30 | 2021-03-30 | 上海傲睿科技有限公司 | Single cell screener, screening assembly, screening method and application |
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| CN112029662A (en) * | 2020-11-06 | 2020-12-04 | 深圳市赛特罗生物医疗技术有限公司 | Cell sorting magnetic grid structure, manufacturing method and magnetic grid tube |
| CN112094839A (en) * | 2020-11-06 | 2020-12-18 | 深圳市赛特罗生物医疗技术有限公司 | Automatic cell magnetic sorting method and device |
| CN112029662B (en) * | 2020-11-06 | 2021-02-05 | 深圳市赛特罗生物医疗技术有限公司 | Cell sorting magnetic grid structure, manufacturing method and magnetic grid tube |
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| CN112553043A (en) * | 2020-12-09 | 2021-03-26 | 深圳先进技术研究院 | Microfluidic chip for separating and purifying fetal nucleated red blood cells |
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