CN107164213A - One kind is based on the cellifugal chip of principle of inertia point - Google Patents
One kind is based on the cellifugal chip of principle of inertia point Download PDFInfo
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- CN107164213A CN107164213A CN201710263764.3A CN201710263764A CN107164213A CN 107164213 A CN107164213 A CN 107164213A CN 201710263764 A CN201710263764 A CN 201710263764A CN 107164213 A CN107164213 A CN 107164213A
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
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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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
Abstract
The invention discloses one kind based on the cellifugal chip of principle of inertia point, runner (3) including matrix and in the matrix, runner (3) is spiral shape or arc, and with the first side wall (S1) and second sidewall (S2) being oppositely arranged；The tongue being spaced apart along the runner (3) is provided with the first side wall (S1), so that the runner (3) formation be interspersed widen section and compression section, the region that tongue is distributed with section correspondence the first side wall is widened, compression section, which is then corresponded on the first side wall, does not have the region of tongue；The runner (3) is used to separate the cell being mixed with the cell solution of sheath fluid stream by size.The present invention is improved by the structure of the runner crucial to its its set-up mode etc., and the problem of both single helical structure and single unilateral reducing and expansion array structure cell separation efficiency are low is can effectively solve the problem that compared with prior art.
Description
Technical field
The invention belongs to micro-fluidic field, more particularly, to one kind based on the cellifugal chip of principle of inertia point.
Background technology
Microfluidic chip technology is a kind of science and technology manipulated in microscale spatial to fluid, utilizes the technology
When separating cell, method is simple, Success rate of virus isolation is high, it is easy to popularizes, receives significant attention.
The cellifugal flow passage structure of foundation principle of inertia point proposed in existing document and patent is generally：Screw type
With reducing and expansion array structure.Both structures have some defects.For spiral-shaped structure, the pressure that can bear due to chip
It is strong limited, so FD (that is, Dien power) and FL (that is, inertia lift) that the structure is provided are smaller, limit separative efficiency.It is right
For reducing and expansion array structure, although it can provide larger FD and FL by reducing and expansion array, but can not be in whole runner
In FD is provided always, only when cell from widen section flow into contraction section when could be influenceed by FD.
By taking helical structure as an example, prior art literature【3】A kind of helical structure based on inertial separation principle is disclosed to use
In isolating red blood cell from blood.The structure is using a sample inlet, a screw-shaped flow paths and a y-bend outlet.Should
Helical flow path is made up of 5 circle Archimedian screw shape microchannels, and the cross section of passage is rectangular surfaces, the rectangular surfaces it is a width of
100um, a height of 50um, curve overall length 13cm, the radius of curvature of spiral most inner side is 3mm.The scholar is by blood with 0.15m/s's
Speed is injected from sample inlet, and 7.32um particles are collected in the lower exit of two prongs, is collected at the upper outlet of two prongs
1.9um particle.
As shown in figure 3, the helical structure realizes that cell separation principle is：Cell can be by inertia lift in bending runner
FL and Dien power two power of FD collective effect.Balance between FL and FD determines equilbrium position of the cell in microchannel.When
During FL >=FD, cell is moved toward side wall S1, as FL ＜ FD, and cell is moved toward side wall S2；FL suffered by larger-size cell compared with
Greatly, FD suffered by the less cell of size is larger.By the runner of certain length, big particle is just separated with small particles.
In the above-mentioned methods, the pressure that can bear due to chip is limited, so FD and FL that the structure is provided are smaller,
Limit separative efficiency.Furthermore, limited by its flow passage structure, the spacing between lateral balancing location residing for different cells compared with
It is small.Finally in order to reach expected separating effect, required runner total length is often longer, has other documents to devise unilateral reducing and expansion
Array structure is used for cell separation, and the runner total length needed for the structure is shorter, but in this design, and cell is only from opening up
Wide section enter compression section when, just can by two power of FD and FL collective effect.Remaining stage is only influenceed by inertia lift FL.
The time that FD is acted on cell is very of short duration, and it cannot be guaranteed that within the so short time whether all cells all by
FD influence.
Labor, prior art literature are carried out below for unilateral reducing and expansion array structure【1】Disclose a kind of unilateral contracting
Expand array structure, the structure is using sample inlet, a sheath liquid flow inlet, two outlets, and chamber and five are widened in six one side
Individual contraction section, to washed corpuscles and blood plasma (size is 2um and below 2um cell)；The scholar is with 1.2mL/h speed
Rate injects blood sample toward sample inlet, injects phosphate buffer toward sheath liquid flow inlet with 12mL/h speed, goes out at two
Haemocyte (size is more than 2um cell) and blood plasma (size is 2um and below 2um cell) are have collected respectively at mouthful,
But blood plasma (size is 2um and below 2um cell) separative efficiency is only 62.2%.
Unilateral reducing and expansion array structure realizes that cell separation principle is：As shown in figure 4, in long and straight type Rectangular Microchannel, carefully
Born of the same parents are mainly influenceed by inertia lift FL, so that cell is focused on close to two equilbrium positions up and down on the long side of rectangle；Curved
In shape runner, cell can be by inertia lift FL and the collective effect of Dien power two power of FD.When liquid is from section is widened into pressure
During contracting section, the path for widening the liquid of section to bend accelerates into compression section, generates Dien vortex, and cell is vortexed in this Dien
It is middle to be influenceed by Dien power FD, so cell is from when widening section into compression section, it will by two power of FL and FD
Collective effect.Location is relevant on the cross section of raceway groove with cell for FL and FD size.Balance between FL and FD
Determine equilbrium position of the cell in microchannel.As FL >=FD, cell is moved toward side wall S1, and as FL ＜ FD, cell is past
Side wall S2 is moved；FL suffered by larger-size cell is larger, and FD suffered by the less cell of size is larger.By several reducing and expansion battle array
Row, haemocyte (size is more than 2um cell) and blood plasma (size is 2um and below 2um cell) are just separated.
In the above-mentioned methods, cell can be just total to only from when widening section into compression section by two power of FD and FL
Same-action.Remaining stage is only influenceed by inertia lift FL.The time that FD is acted on cell is very of short duration, and it cannot be guaranteed that
Whether all cells all receive FD influence in the so short time.
Secondly as in other stages, cell is only influenceed by FL, the cell that FL can will move to side wall S1 and S2 is pulled to
Runner center so that the cell in expectational equilibrium position deviates equilbrium position, so as to reduce separation to a certain extent
Efficiency.
Bibliography is as follows：
【1】"Inertial blood plasma separation in a contraction–expansion array
microchannel"；
【2】"Enhanced blood plasma separation by modulation of inertial lift
force"；
【3】"Continuous particle separation in spiral microchannels using dean
flows and differential migration"；
【4】"Improved understanding of particle migration modes in spiral
inertial microfluidic devices"；
【5】"Continuous inertial microparticle and blood cell separation in
straight channels with local microstructures"。
The content of the invention
For the disadvantages described above or Improvement requirement of prior art, principle of inertia is based on it is an object of the invention to provide one kind
Point cellifugal chip, wherein by set-up mode of flow passage structure crucial to its etc. (especially to widened in runner section with
The shape and relevant parameter of compression section) it is improved, single helical structure, Yi Jidan are can effectively solve the problem that compared with prior art
The problem of both one unilateral reducing and expansion array structure cell separation efficiency are low, and the present invention widens section by controlling in runner every section
With the length of compression section, width and runner overall length and height so that the chip especially can using 3um and 6um sizes as boundary,
Separate cell.
To achieve the above object, it is proposed, according to the invention, there is provided one kind based on the cellifugal chip of principle of inertia point, its feature
It is, including matrix and the runner (3) in the matrix, the sample that the chip also includes being connected with the runner (3) enters
Mouth (1), sheath liquid flow inlet (2), large scale cell outlet (4) and small size cell outlet (5)；
Wherein,
The sample inlet (1) is used for the cell solution that processing to be separated is inputted to the entrance of the runner (3)；
The sheath liquid flow inlet (2) be used for the runner (3) entrance input sheath fluid stream, the sheath fluid stream be used for it is described
Cell solution is mixed；
The runner (3) is spiral shape or arc；The runner (3) has the first side wall (S1) being oppositely arranged and the second side
Wall (S2), wherein the first side wall (S1) is located at the inner side close to spiral center or arc center, the second sidewall (S2)
It is located remotely from the outside at spiral center or arc center；It is provided with along the runner (3) and is spaced apart on the first side wall (S1)
Tongue so that what the runner (3) formation was interspersed widens section and compression section, and described widen is distributed on section correspondence the first side wall
There is the region of tongue, the compression section, which is then corresponded on the first side wall, does not have the region of tongue；The runner (3) is used for being mixed with sheath
Cell in the cell solution of liquid stream is separated by size；
The large scale cell outlet (4) is located at close to the side at spiral center or arc center, for from the runner
(3) cell separation liquid of the outlet output containing large scale cell；
The small size cell outlet (5) is located remotely from the side at spiral center or arc center, for from the runner
(3) cell separation liquid of the outlet output containing small size cell；
In addition, the entrance and exit of the runner (3) is located at the two ends of the runner (3) respectively.
As present invention further optimization, in the runner (3), widened described in any one section the arc length of section for 300~
700um, the arc length of any one section of compression section is 300~1200um；It is preferred that, the width for widening section is 350um, institute
The width for stating compression section is 50um；The runner of the runner (3) is highly 20~25um；It is preferred that, second sidewall on the runner (3)
(S2) radius of curvature near the spiral center or the arc-shaped center section is 5mm~7mm.
As present invention further optimization, in the runner (3), the arc length that section is widened described in any one section is 700um,
The arc length of any one section of compression section is 1200um；The height of the runner (3) is 25um, and the overall length of the runner (3) is
23mm, the number of required compression section is 11, and the number for widening section is that second sidewall (S2) is most on 12, the runner (3)
Radius of curvature close to the spiral center or the arc-shaped center section is 7mm, and the particle diameter of the large scale cell is not less than
6um, the particle diameter of the small size cell is no more than 3um.
As present invention further optimization, the cell of the processing to be separated inputted from the sample inlet (1) is molten
The flow velocity of liquid is 1 with the ratio between the flow velocity of the sheath fluid stream inputted from the sheath liquid flow inlet (2)：5.
As present invention further optimization, when the runner (3) is spiral shape, the runner (3) preferably presses A Ji meter
Moral helical is distributed.
As present invention further optimization, described matrix includes the upper matrix and lower substrate being superimposed up and down, the runner
(3) it is located between the upper matrix and the lower substrate.
As present invention further optimization, the sheath fluid stream is phosphate buffer.
It is used as present invention further optimization, the rectangular cross-section of the runner (3).
Based on the cellifugal chip of principle of inertia point by the way that runner is arranged to unilateral reducing and expansion array in the present invention
Spiral/arcuate structure, can effectively improve cell separation efficiency.The present invention is close to runner spiral center or arc center
The tongue being spaced apart along the runner is set on madial wall, is formed and widens section and the stream of compression segment structure with what is be interspersed
Road, has further excavated the excellent of reducing and expansion array structure and helical flow path structure (or arced flow path structure) both flow passage structures
Gesture, makes full use of the characteristics of Dien power is with inertia lift in runner different zones so that reducing and expansion array structure and helical flow path knot
Both structures (or arced flow path structure) are organically combined, and effectively raise cell separation efficiency.
The present invention is to set along the runner to be spaced apart on the madial wall close to runner spiral center or arc center
Tongue, can not reach a point cellifugal purpose if the tongue being spaced apart along the runner is set along along lateral wall.When along outside
When wall sets tongue, the inertia lift suffered by bulky grain is to point to side wall S2 by side wall S1, and inertia lift now can be by big grain
Son is dragged to side wall S2；At the same time, the raceway groove small particles are when widening section into compression section, and the Dien power received is also from side wall S1
Side wall S2 is pointed to, so that little particle is also dragged to side wall S2 from side wall S1, does not reach expected separating effect.
Table 1：3um is with 6um particles in the unilateral scalable array circular arc of inside curved unilateral scalable array arc structure and bent outward
Structure standoff distance
Note：Standoff distance now is the standoff distance of 6um particles bottom and 3um particles the top in exit, this
The radius of curvature of circular arc lateral wall is 7mm.Described in table 6 widen that segment table shows is that the raceway groove has 6 to widen section and 5
Compression section (general few one) of quantity than widening section of the quantity of compression section.
Radius of curvature of the invention near the spiral center or the arc-shaped center section is 5mm~7mm.When this
When the radius of curvature of invention is less than 5mm, it can cause to accommodate not lower inlet 1 in the most raceway groove of inner side, entrance 2 is closed with regard to structure design
For rationality, radius of curvature of the invention is not less than 5mm.When radius of curvature is more than 7mm, the separation effect of the present invention can be reduced
Rate.
Table 2：3um and separating effect of the 6um particles under different curvature radius
The height of the structure of the present invention is general in 20~25um.When separating 2um and more than 2um particle, this structure
Height is 25um.When separating below 2um particle, the height of this structure is 20um.To ensure that particle can be made in inertia lift
The single-row particle beams, particle diameter a are formed with lower focusingpIt is that runner height h needs to meet a with flow path features sizep/h≥0.07。
Because bulky grain is mainly influenceed by inertia lift in channels, want to allow bulky grain to flow from outlet in straight line in channels
Go out, it is necessary to meet ap/h≥0.07.Therefore when separation particle size becomes small, channel height also should become short therewith.
Table 3：3um and standoff distance of the 6um particles under different height
Note：The radius of curvature of this circular arc lateral wall is 7mm.
Relative to single helical structure, the unilateral reducing and expansion array helical structure in the present invention can be in shorter runner
Separate cell.When cell movement is to when widening flow channel section, runner is widened so that the wall induction inertia lift that cell is subject to is dashed forward
So reduce, small size cell can be moved to S2 sides wall, and stably in new equilbrium position, large scale cell can be transported to S1 sides wall
It is dynamic, and stably caused in new equilbrium position between the equilbrium position of large scale cell and the cell balance position of small size
Spacing becomes big.When liquid is from when widening section into compression section, the path for widening the liquid of section to bend accelerates into compression section, should
Acceleration can be such that FL and FD suffered by cell strengthens to a certain extent, so that cell can faster be separated.
Based on principle of inertia point cellifugal chip in the present invention, its unilateral reducing and expansion array spiral flow channel (or unilateral contracting
Expand array arced flow path), cell can be allowed all to be influenceed in whole runner by Dien power, it is former based on inertia relative to existing
The cellifugal reducing and expansion array structure of reason point, can effectively overcome existing reducing and expansion array structure only to work as cell and flow into receipts from section is widened
This defect could be influenceed during contracting section by Dien power.
In the existing point cellifugal reducing and expansion array structure based on principle of inertia, the cell in compression section postmedian position only by
The influence of inertia lift, particle can be focused on close to long paracentral two balances as shown in Figure 4 under the effect of pure inertia lift
On position, therefore the cell in compression section postmedian position can deviate expected focusing equilbrium position to runner off-centring；And this
The runner with unilateral reducing and expansion array helical structure (or arcuate structure) employed in invention, can allow cell entirely flowing
The coupling of Dien power and inertia lift is constantly subjected in road, so that cell is fixed on expected equilbrium position.
The present invention improves the separation effect of helical structure to a certain extent using unilateral reducing and expansion array spiral flow channel
Rate.For helical structure, as shown in Figure 5 when cell is in equilbrium position, the lateral balancing location residing for different cells
Between spacing it is smaller, easily cause part mixing with cells, cause separative efficiency to reduce.The unilateral reducing and expansion array junctions that the present invention is designed
Special unilateral reducing and expansion array can allow small size cell to be moved to S2 sides wall in structure spiral flow channel, and large scale cell can be to S1
Side wall motion, causes the spacing between the equilbrium position of large scale cell and the cell balance position of small size to become big, so as to carry
High separating efficiency.
For existing single helical structure, to reach expected separating effect, required runner total length is general longer.
In the case that separating effect is consistent, the runner designed by the present invention can shorten runner total length to a certain extent.
For single helical structure, because the channel dimensions are longer, the pressure that chip can bear is limited, in a constant current
In fast, the FD and FL that the structure is provided are smaller, limit separative efficiency.The reducing and expansion array that the present invention is designed can solve this and lack
Fall into, when cell is from when widening section and flowing into compression section, runner diminishes suddenly so that fluid accelerates into compression section, and now fluid
Speed in several times growth, flow velocity speed offer one larger FD and FL can be provided so that cell can reach faster
Equilbrium position.
In addition, being had further the advantage that in the present invention based on the cellifugal chip of principle of inertia point：
(1), it is miniaturized.Whole chip area is only several square centimeters.Required reagent volume is only a microlitre rank.
(2) separative efficiency, is improved.Cell of different sizes is caused to pass through several circular arcs using above-mentioned micro-fluidic chip
Shape compression is widened array and can just separated.
(3), online observation.The micro-fluidic chip can be observed directly under CCD inverted microscopes, use high-speed camera
Machine carries out image record, easy to operate.
(4) it is, cheap.Chip material can use PDMS and lucite.Reagent dosage is few, it will be apparent that reduce examination
Agent cost.
Brief description of the drawings
Fig. 1 is its overall structure diagram of separation cell chip and the office that the present invention has unilateral reducing and expansion array helical structure
Portion's amplification signal；
Fig. 2 is its overall structure diagram of separation cell chip and the office that the present invention has unilateral reducing and expansion array arc structure
Portion's amplification signal；
Fig. 3 is that the chip of helical structure in the prior art realizes the principle schematic of cell separation；
Fig. 4 be in the prior art single particle in straight channel only by present position schematic diagram during inertia lift；
Fig. 5 is that the chip of helical structure in the prior art realizes the schematic diagram of cell separation；
Fig. 6 A are distributed simulation figure of the 6um particle in the arc-shaped structure along outer wall tongue, and Fig. 6 B are figures
6A partial enlarged drawing；
Fig. 7 A are distributed simulation figure of the 3um particle in the arc-shaped structure along outer wall tongue, and Fig. 7 B are figures
7A partial enlarged drawing.
The implication of each reference is as follows in figure：1 is sample inlet, and 2 be sheath liquid flow inlet, and 3 be cell separation unit
(that is, runner), 4 be large scale cell outlet, and 5 be small size cell outlet, and S1 is madial wall (that is, the first side wall), and S2 is outer
Side wall (that is, second sidewall).
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, it is right below in conjunction with drawings and Examples
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.As long as in addition, technical characteristic involved in each embodiment of invention described below
Not constituting conflict each other can just be mutually combined.
The invention discloses one kind based on the cellifugal micro-fluidic chip of principle of inertia point, the micro-fluidic chip is from top to bottom
Including upper matrix, lower substrate, the upper matrix can be fixedly connected by the method and lower substrate of bonding；Runner tongue is located at
(projection of the runner tongue on chip plane is as shown in Figure 1 in upper substrate；Lower substrate with can on the surface of upper substrate contact
To be not provided with any tongue, at the same lower substrate with the surface of upper substrate contact parallel to chip plane), as shown in figure 1, described
Flow channel layer is a unilateral reducing and expansion array structure spiral flow channel, including sample inlet 1, sheath liquid flow inlet 2, a big chi
Very little cell outlet 4 and small size cell outlet 5.Cell can be made in runner by Dien power FD and inertia lift FL coupling
With after the runner for flowing through certain length, various sizes of cell focuses on different lateral balancing locations, is finally recovered out not
With the cell of size.This chip can realize cell separation in the case where flow velocity is higher, and required flow passage structure is simple, operation
Simply, flux is high, and improves cell separation efficiency to a certain extent.
It is kind of the cell separation core with novel flow channel structure based on the cellifugal chip of principle of inertia point in the present invention
Piece, can efficiently, it is easy, quickly separate cell, the method for preparation and use of the chip can be carried out using following steps：
1st, flow passage structure is designed.The runner, which is one section, has the helical flow path of unilateral reducing and expansion array structure, and runner one end is
The sample and sheath liquid flow inlet of one Y-shaped, 1 is sample inlet, and 2 be sheath liquid flow inlet, and the other end is the big cellule of a Y-shaped
Outlet, 3 be the outlet of large scale cell, and 4 be the outlet of small size cell.Described microchannel cross section is rectangle.
2nd, channel size is designed.In the runner reducing and expansion array, section arc length is widened for 300~700um, width is 350um, pressure
Contracting section arc length is 300~1200um, and width is 50um, and runner is highly 20~25um.
3rd, chip is made.The chip includes upper matrix and lower substrate, and the material of upper matrix can be dimethyl silicone polymer
(PDMS), lower substrate material can be glass.Two matrixes are combined together by way of bonding.Runner tongue is located at upper base
In body.
4th, cell solution is handled.The cell sample diluted to a certain degree in this scenario can greatly reduce the wind of runner blocking
Danger.
5th, in the technical scheme, micro-fluidic chip sample introduction needs dynamical system.Using dynamical system by cell sample and
Sheath fluid stream injects from injection port 1 and injection port 2 simultaneously at the appropriate speed.
6th, observation experiment result.
It is specific embodiment below：
Embodiment 1
With reference to shown in Fig. 2, the cell separation chip includes the microchannel on matrix and matrix, and the passage includes cell
Sample feeding mouthful 1, sheath fluid stream injection port 2, cell separation unit 3, large scale cell outlet 4 and small size cell outlet 5.
In the technical scheme, micro-fluidic chip sample introduction needs dynamical system, dynamical system be used for by cell solution and sheath fluid stream respectively from
Injection port 1 and injection port 2 are continuously injected into chip, and the dynamical system in this example is syringe pump.Sheath fluid stream is used to enter
Section is widened for first after mouthful to be focused into a branch of by cell and press close to S1 sides wall and enter compression section.
Cell separation unit 3 is circular arc passage, and the passage is inwardly curved from cell injection port 1.Lateral wall S2 is inflexed
Radius of curvature is 7mm, and the size of the passage is that any section arc length of widening of runner is 700um, and width is 350um, any compression
Section arc length is 1200um, and width is 50um, and whole runner is highly 25um, and the length 23mm of cell separation unit 3 (further comprises
The length of intake section and exit portion).Cell injection port 1, sheath fluid stream injection port 2, large scale particle outlet 4 diameter can be excellent
Elect 400um as, and the diameter of small size cell outlet 5 is preferably 400um.
With reference to shown in Fig. 2, the cell principle using the runner separation different size is：In the microchannel of continuous bend,
Cell is mainly by two kinds of power of lift FL and Dien power FD.Balance between FL and FD determines balance position of the cell in microchannel
Put.As FL >=FD, cell is moved toward side wall S1, as FL ＜ FD, and cell is moved toward side wall S2；Larger-size cell institute
Larger by FL, FD suffered by the less cell of size is larger.When cell movement is to when widening flow channel section, runner is widened so that cell
The wall induction inertia lift being subject to reduces suddenly, and small size cell can be moved to S2 sides wall, and stably in new equilbrium position,
Large scale cell can be moved to S1 sides wall, and stably cause the equilbrium position of large scale cell and small chi in new equilbrium position
Spacing between very little cell balance position becomes big.When liquid is from when widening section into compression section, widen the liquid of section to bend
Path accelerate into compression section, the acceleration can be such that FL and FD suffered by cell strengthens to a certain extent, so as to allow cell energy
It is enough faster to separate.
The flow velocity of two samples can influence cell separation effect, the separation produced by different velocity ratios in the technical scheme
Efficiency is also different.The speed ratio of cell sample and sheath fluid stream is 1 in this example:5.Using two syringe pumps by cell sample
(blood of dilution) and sheath fluid stream (phosphate buffer) are respectively with 0.012m/s, 0.06m/s speed simultaneously from the and of injection port 1
Injection port 2 injects.(it is 0.8m/s now to correspond to maximal rate in raceway groove)
During detection, observed using CCD inverted microscopes, videograph is carried out with high-speed camera., can by observation
To see that haemocyte (cell of the size as more than 6um) is gradually moved toward side wall S1, (size is thin for 3um's and below 3um for blood plasma
Born of the same parents) gradually moved toward side wall S2.
Micro-fluidic chip workflow includes：
1st, two syringe pumps are got out, syringe is extracted to the blood cell and phosphate buffer diluted respectively,
It is placed on syringe pump and is connected by teflon pipe with chip.
2nd, chip is placed on correct position under CCD inverted microscopes, focused, clearly observed.
3rd, syringe pump is opened, flow velocity is adjusted, the various sizes of cell separation of observation is realized.
Table 4：Under friction speed, the distance that 3um is separated with 6um particles in straight flute road with unilateral scalable array arc structure
Note：Standoff distance now is 6um particles bottom and 3um particles topmost in the standoff distance in exit.This
The radius of curvature of arc-shaped structure is 7mm.Described in table 6 widen that segment table shows is that the raceway groove has 6 to widen section and 5 pressures
Contracting section.Described in table 12 widen that segment table shows is that the raceway groove has 12 to widen section and 11 compression sections.
Table 5：The distance that 2um is separated with 7um particles in unilateral scalable arc-shaped structure with helical structure
Note：Standoff distance now is the standoff distance of 7um particles bottom and 2um particles the top in exit, this
The radius of curvature of arc-shaped structure is 7mm.The helical structure is Continuous particle separation in
Spiral microchannels using dean flows and differential migration are document【3】In
Structure, the structural model a height of 50um, a width of 100um, most inner side radius of curvature is 3mm, and overall length is 13cm., the speed in the table
Spend speed when optimal for correspondence model separation efficiency.
Table 6：The distance that 3um is separated from 7um particles in three kinds of different models
Note：Standoff distance now is 6um particles bottom and 3um particles topmost in the standoff distance in exit.Two
The radius of curvature of individual arc-shaped structure is 7mm.The size of arc structure is：Wide 50um, arc length 21.6mm.Now both are justified
The arc length of arc is equal with the length of straight channel.
As seen from the above table, when secondary three kinds of moulded dimensions are consistent, when speed is consistent, 3um of the present invention and 7um particle phase gauges
From for 141.6mm, more than other two structure separating distance sum.
In the present invention, entrance (including sample inlet 1, sheath liquid flow inlet 2) and outlet (including it is large scale cell outlet 4, small
Size cell outlet 5) position can exchange, now, runner is widened the corresponding tongue of section and is still located on madial wall S1, large scale
Cell outlet 4 is still located on madial wall S1 the same side, and small size cell outlet 5 is still located on lateral wall S2 the same side.
Runner in the present invention both can be spiral shape or arc, can be according to specific cell granulations to be separated
Depending on size, for example, can see by table 4, when unilateral scalable helical structure is longer, the optimal velocity needed for separate particles is just
Smaller, the optimal velocity as needed for 6 are widened section unilateral scalable array arc structure separation 3um and 6um particles is 1.3m/s, and 12
The individual unilateral scalable array arc structure separation 3um of section and optimal velocity needed for 6um particles of widening is 0.8m/s.Due to chip size
It is smaller, and be to be combined together upper lower substrate by the mode of key sum, therefore the pressure that core is born is limited, when optimal velocity reduction
When, can effectively it reduce because raceway groove inside water pressure is excessive, the probability for causing liquid to be oozed out from porch.In actual use,
If when less high to rate request, longer raceway groove can be used, to reduce because pressure is excessive in chip, and caused brokenly
The probability for splitting and seeping water.
The arc of runner in the present invention can be elliptic arc or positive circular arc；When arc is circular arc, arc center is
The circular arc center of circle.In addition, the runner height in the present invention is adjusted flexibly also according to the diameter of particle to be separated, for example, separation 10um
Height can be used to be 25um for particle to 2um, and height can be used to be 20um for separation 1.7um to 0.5um particle.
The radius of curvature of runner is on the basis of runner second sidewall S2 radius of curvature in the present invention；When runner is circle
Arc, then the radius of curvature holding of runner everywhere is constant；When runner is spiral shape, second sidewall S2 is near spiral center portion
The radius of curvature divided is 5mm~7mm (corresponding to runner close to one end of spiral center).
As it will be easily appreciated by one skilled in the art that the foregoing is merely illustrative of the preferred embodiments of the present invention, it is not used to
The limitation present invention, any modifications, equivalent substitutions and improvements made within the spirit and principles of the invention etc., it all should include
Within protection scope of the present invention.
Claims (8)
1. one kind is based on the cellifugal chip of principle of inertia point, it is characterised in that the stream including matrix and in the matrix
Road (3), the chip also includes sample inlet (1), sheath liquid flow inlet (2), the large scale cell outlet being connected with the runner (3)
And small size cell outlet (5) (4)；Wherein,
The sample inlet (1) is used for the cell solution that processing to be separated is inputted to the entrance of the runner (3)；
The sheath liquid flow inlet (2) is used to input sheath fluid stream to the entrance of the runner (3), and the sheath fluid stream is used for and the cell
Solution is mixed；
The runner (3) is spiral shape or arc；The runner (3) has the first side wall (S1) being oppositely arranged and second sidewall
(S2), wherein the first side wall (S1) is located at the inner side close to spiral center or arc center, second sidewall (S2) position
In the outside away from spiral center or arc center；It is provided with what is be spaced apart along the runner (3) on the first side wall (S1)
Tongue so that what the runner (3) formation was interspersed widens section and compression section, described widen is distributed with section correspondence the first side wall
The region of tongue, the compression section, which is then corresponded on the first side wall, does not have the region of tongue；The runner (3) is used for being mixed with sheath fluid
Cell in the cell solution of stream is separated by size；
The large scale cell outlet (4) is located at close to the side at spiral center or arc center, for from the runner (3)
Cell separation liquid of the outlet output containing large scale cell；
The small size cell outlet (5) is located remotely from the side at spiral center or arc center, for from the runner (3)
Cell separation liquid of the outlet output containing small size cell；
In addition, the entrance and exit of the runner (3) is located at the two ends of the runner (3) respectively.
2. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that in the runner (3), arbitrarily
The arc length that section is widened described in one section is 300~700um, and the arc length of any one section of compression section is 300~1200um；It is preferred that
, the width for widening section is 350um, and the width of the compression section is 50um；The runner of the runner (3) highly for 20~
25um；It is preferred that, curvature of the second sidewall (S2) near the spiral center or the arc-shaped center section on the runner (3)
Radius is 5mm~7mm.
3. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that in the runner (3), arbitrarily
The arc length that section is widened described in one section is 700um, and the arc length of any one section of compression section is 1200um；The height of the runner (3)
Spend for 25um, the overall length of the runner (3) is 23mm, and the number of required compression section is 11, and the number for widening section is 12,
On the runner (3) second sidewall (S2) near the spiral center or the arc-shaped center section radius of curvature be 7mm, institute
The particle diameter for stating large scale cell is not less than 6um, and the particle diameter of the small size cell is no more than 3um.
4. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that from the sample inlet (1)
The flow velocity of the cell solution of the processing to be separated of input and the stream of the sheath fluid stream inputted from the sheath liquid flow inlet (2)
The ratio between speed is 1：5.
5. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that when the runner (3) is spiral shell
When revolving shape, the runner (3) is preferably distributed by Archimedes spiral.
6. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that described matrix includes folded up and down
Plus upper matrix and lower substrate, the runner (3) be located between the upper matrix and the lower substrate.
7. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that the sheath fluid stream is phosphate
Buffer solution.
8. as claimed in claim 1 based on the cellifugal chip of principle of inertia point, it is characterised in that the section of the runner (3)
For rectangle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710263764.3A CN107164213A (en) | 2017-04-21 | 2017-04-21 | One kind is based on the cellifugal chip of principle of inertia point |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710263764.3A CN107164213A (en) | 2017-04-21 | 2017-04-21 | One kind is based on the cellifugal chip of principle of inertia point |
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| CN107159326A (en) * | 2017-05-09 | 2017-09-15 | 西北工业大学 | Inertia based on enhanced Secondary Flow focuses on chip |
| CN108246374A (en) * | 2018-03-28 | 2018-07-06 | 东南大学 | Three-dimensional spiral Constituent cell sorting micro-fluidic chip and preparation method thereof |
| CN108715794A (en) * | 2018-05-08 | 2018-10-30 | 南京师范大学 | A kind of cell accurately manipulates micro-fluidic device |
| CN109666584A (en) * | 2018-12-29 | 2019-04-23 | 北京工业大学 | A kind of experimental provision can be used for carrying out circulating tumor cell sorting experiment |
| CN109967150A (en) * | 2019-04-24 | 2019-07-05 | 河海大学常州校区 | It is a kind of for manipulating the inertia micro-fluidic chip of micro-nano granules |
| CN110124757A (en) * | 2019-04-29 | 2019-08-16 | 南京师范大学 | A kind of micro particles sorting micro-fluidic device |
| CN111139182A (en) * | 2018-11-02 | 2020-05-12 | 青岛华大智造普惠科技有限公司 | Magnetic screening device, micro-droplet screening system and magnetic screening method of micro-droplets |
| CN111690508A (en) * | 2020-05-19 | 2020-09-22 | 东南大学 | Multi-functional unit integrated rare tumor cell multi-stage sorting device |
| CN111909823A (en) * | 2019-05-08 | 2020-11-10 | 清华大学 | Inertial micro-fluidic chip for enriching circulating tumor cells |
| CN112007704A (en) * | 2020-07-08 | 2020-12-01 | 河海大学常州校区 | Micro-fluidic chip and method for sorting micro-nano particles by inertial turbulence |
| CN112354573A (en) * | 2020-10-26 | 2021-02-12 | 深圳亘流科技有限公司 | Step type inertial focusing micro-fluidic chip |
| CN112827521A (en) * | 2021-02-24 | 2021-05-25 | 南京航空航天大学苏州研究院 | Inertial separation microfluidic chip based on electrical impedance imaging visual detection |
| CN114260036A (en) * | 2021-12-23 | 2022-04-01 | 西安交通大学 | Micro-fluidic chip based on inertial focusing sorting |
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| CN107159326A (en) * | 2017-05-09 | 2017-09-15 | 西北工业大学 | Inertia based on enhanced Secondary Flow focuses on chip |
| CN108246374A (en) * | 2018-03-28 | 2018-07-06 | 东南大学 | Three-dimensional spiral Constituent cell sorting micro-fluidic chip and preparation method thereof |
| CN108246374B (en) * | 2018-03-28 | 2020-09-11 | 东南大学 | Three-dimensional spiral structure cell sorting micro-fluidic chip and manufacturing method thereof |
| CN108715794A (en) * | 2018-05-08 | 2018-10-30 | 南京师范大学 | A kind of cell accurately manipulates micro-fluidic device |
| CN111139182A (en) * | 2018-11-02 | 2020-05-12 | 青岛华大智造普惠科技有限公司 | Magnetic screening device, micro-droplet screening system and magnetic screening method of micro-droplets |
| CN111139182B (en) * | 2018-11-02 | 2023-11-14 | 青岛华大智造科技有限责任公司 | Magnetic screening device, micro-droplet screening system and micro-droplet magnetic screening method |
| CN109666584A (en) * | 2018-12-29 | 2019-04-23 | 北京工业大学 | A kind of experimental provision can be used for carrying out circulating tumor cell sorting experiment |
| CN109967150A (en) * | 2019-04-24 | 2019-07-05 | 河海大学常州校区 | It is a kind of for manipulating the inertia micro-fluidic chip of micro-nano granules |
| CN110124757A (en) * | 2019-04-29 | 2019-08-16 | 南京师范大学 | A kind of micro particles sorting micro-fluidic device |
| CN111909823B (en) * | 2019-05-08 | 2023-04-18 | 清华大学 | Inertial micro-fluidic chip for enriching circulating tumor cells |
| CN111909823A (en) * | 2019-05-08 | 2020-11-10 | 清华大学 | Inertial micro-fluidic chip for enriching circulating tumor cells |
| CN111690508A (en) * | 2020-05-19 | 2020-09-22 | 东南大学 | Multi-functional unit integrated rare tumor cell multi-stage sorting device |
| CN112007704A (en) * | 2020-07-08 | 2020-12-01 | 河海大学常州校区 | Micro-fluidic chip and method for sorting micro-nano particles by inertial turbulence |
| CN112354573B (en) * | 2020-10-26 | 2022-01-04 | 深圳亘流科技有限公司 | Step type inertial focusing micro-fluidic chip |
| CN112354573A (en) * | 2020-10-26 | 2021-02-12 | 深圳亘流科技有限公司 | Step type inertial focusing micro-fluidic chip |
| CN112827521A (en) * | 2021-02-24 | 2021-05-25 | 南京航空航天大学苏州研究院 | Inertial separation microfluidic chip based on electrical impedance imaging visual detection |
| CN114260036A (en) * | 2021-12-23 | 2022-04-01 | 西安交通大学 | Micro-fluidic chip based on inertial focusing sorting |
| CN114260036B (en) * | 2021-12-23 | 2023-01-31 | 西安交通大学 | Micro-fluidic chip based on inertial focusing sorting |
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Application publication date: 20170915 |