CN106281965A - Large scale network array unicellular capture micro-fluidic device - Google Patents
Large scale network array unicellular capture micro-fluidic device Download PDFInfo
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- CN106281965A CN106281965A CN201610671600.XA CN201610671600A CN106281965A CN 106281965 A CN106281965 A CN 106281965A CN 201610671600 A CN201610671600 A CN 201610671600A CN 106281965 A CN106281965 A CN 106281965A
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- capture
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
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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 a kind of large scale network array unicellular capture micro-fluidic device, including cell capture cavity, it is provided with inflow entrance and flow export on described cell capture cavity, it is provided with cell capture device in described cell capture cavity, described cell capture device includes multiple unicellular capturing unit, the plurality of unicellular capturing unit is connected with each other composition preset shape according to preset structure, each described unicellular capturing unit includes capturing runner and bypass runner, it is provided with slot on described capture runner, described slot matches with the size of cell to be captured, the capture runner of each described unicellular capturing unit all connects with capture runner and the bypass runner of adjacent unicellular capturing unit with bypass runner.Present invention have the advantage that unicellular capturing unit presents network-like distribution, it is achieved multi-path structure during capture, and there is adaptivity, capture rate can be improved, reduce the risk of device blocking simultaneously.
Description
Technical field
The present invention relates to cell capture technical field, the specific design one unicellular capture of large scale network array is micro-fluidic
Device.
Background technology
At present, the unicellular capture device in correlation technique is by arranging that substantial amounts of capturing unit, in runner, or makes
By the mode of multi-channel parallel, to improve flux.After cell suspending liquid injects, cell enters catching of design in a random way
Obtaining unit, a small amount of cell is limited in runner by the groove in slot or runner, and other most cells are with suspension flow
Outlet.But the probability that can cell enter capturing unit in runner presents a kind of Gauss distribution, therefore reduces this type of
The capturing efficiency of mode, needs high-throughout cell to flow into the cell capture rate of each unit of guarantee, additionally most devices
Have employed the mode connected in series of capturing unit, cause because of blocking cause device fault, can not reusable risk.
Summary of the invention
It is contemplated that at least solve one of above-mentioned technical problem.
To this end, it is an object of the invention to propose a kind of large scale network array unicellular capture micro-fluidic device, it is achieved
Multi-path structure during capture, and there is adaptivity, capture rate can be improved, reduce the risk of device blocking simultaneously.
To achieve these goals, embodiment of the invention discloses that a kind of large scale network array unicellular capture miniflow
Control device, including cell capture cavity, described cell capture cavity is provided with inflow entrance and flow export, described cell capture chamber
The internal cell capture device that is provided with, described cell capture device includes multiple unicellular capturing unit, the plurality of unicellular
Capturing unit is connected with each other composition preset shape according to preset structure, each described unicellular capturing unit include capture runner and
Bypass runner, described capture runner is provided with slot, and described slot matches with the size of cell to be captured, each described list
Capture runner and the bypass runner of cell capture unit all connect with capture runner and the bypass runner of adjacent unicellular capturing unit
Logical.
Large scale network array unicellular capture micro-fluidic device according to embodiments of the present invention, unicellular capturing unit in
Existing network-like distribution, it is achieved multi-path structure during capture, and there is adaptivity, capture rate can be improved, reduce device simultaneously
The risk of part blocking.
It addition, large scale network array according to the above embodiment of the present invention unicellular capture micro-fluidic device, it is also possible to
There is following additional technical characteristic:
Further, described unicellular capturing unit includes a described capture runner and two described bypass runners, two
Individual described bypass runner constitutes a rectangular configuration, one group of diagonal angle of rectangular configuration described in described capture flow passage.
Further, described cell capture device is matrix structure, and multiple described unicellular capturing units are separately positioned on
On the different element positions of described matrix structure.
Further, when described unicellular capturing unit does not captures cell, the volume flow rate of described capture runner is more than
The volume flow rate of described bypass runner.
Further, being provided with negative pressure and provide device at described flow export, described negative pressure provides device for described
Inflow entrance, between described cell capture device and described flow export formed negative pressure.
Further, described negative pressure provides device to include polyethylene tube and syringe, by syringe described in pull in institute
State inflow entrance, between described cell capture device and described flow export, form negative pressure.
The additional aspect of the present invention and advantage will part be given in the following description, and part will become from the following description
Obtain substantially, or recognized by the practice of the present invention.
Accompanying drawing explanation
Above-mentioned and/or the additional aspect of the present invention and advantage are from combining the accompanying drawings below description to embodiment and will become
Substantially with easy to understand, wherein:
Fig. 1 (A) is that the master of the large scale network array unicellular capture micro-fluidic device of one embodiment of the invention regards knot
Structure schematic diagram；
Fig. 1 (B) is the side-looking knot of the large scale network array unicellular capture micro-fluidic device of one embodiment of the invention
Structure schematic diagram；
Fig. 2 is the principle schematic of the cell capture device capture cell of one embodiment of the invention；
Fig. 3 is the schematic diagram of the theoretical model structure of one embodiment of the invention；
Fig. 4 is the unicellular capture micro-fluidic device acquisition procedure schematic diagram of one embodiment of the invention；
Fig. 5 is 5 × 5 a kind of possible cell capture sequential schematic of cell capture device of one embodiment of the invention；
Fig. 6 (A) is the capturing unit network simulation analysis chart of one embodiment of the invention the first special cells array pattern；
Fig. 6 (B) is the capturing unit network simulation analysis chart of one embodiment of the invention the second special cells array pattern；
Fig. 6 (C) is Fig. 6 (A) experimental result picture；
Fig. 6 (D) is Fig. 6 (B) experimental result picture.
Detailed description of the invention
Embodiments of the invention are described below in detail, and the example of described embodiment is shown in the drawings, the most from start to finish
Same or similar label represents same or similar element or has the element of same or like function.Below with reference to attached
The embodiment that figure describes is exemplary, is only used for explaining the present invention, and is not considered as limiting the invention.
In describing the invention, it is to be understood that term " " center ", " longitudinally ", " laterally ", " on ", D score,
Orientation or the position relationship of the instruction such as "front", "rear", "left", "right", " vertically ", " level ", " top ", " end ", " interior ", " outward " are
Based on orientation shown in the drawings or position relationship, it is for only for ease of the description present invention and simplifies description rather than instruction or dark
The device or the element that show indication must have specific orientation, with specific azimuth configuration and operation, therefore it is not intended that right
The restriction of the present invention.Additionally, term " first ", " second " are only used for describing purpose, and it is not intended that instruction or hint relatively
Importance.
In describing the invention, it should be noted that unless otherwise clearly defined and limited, term " is installed ", " phase
Even ", " connection " should be interpreted broadly, for example, it may be fixing connection, it is also possible to be to removably connect, or be integrally connected；Can
To be mechanical connection, it is also possible to be electrical connection；Can be to be joined directly together, it is also possible to be indirectly connected to by intermediary, Ke Yishi
The connection of two element internals.For the ordinary skill in the art, can understand that above-mentioned term is at this with concrete condition
Concrete meaning in invention.
With reference to explained below and accompanying drawing, it will be clear that these and other aspects of embodiments of the invention.Describe at these
With in accompanying drawing, specifically disclose some particular implementation in embodiments of the invention, represent the enforcement implementing the present invention
Some modes of the principle of example, but it is to be understood that the scope of embodiments of the invention is not limited.On the contrary, the present invention
All changes, amendment and equivalent in the range of spirit that embodiment includes falling into attached claims and intension.
Below in conjunction with accompanying drawing, large scale network array unicellular capture micro-fluidic device according to embodiments of the present invention is described.
Fig. 1 (A) is that the master of the large scale network array unicellular capture micro-fluidic device of one embodiment of the invention regards knot
Structure schematic diagram, Fig. 1 (B) is the side-looking knot of the large scale network array unicellular capture micro-fluidic device of one embodiment of the invention
Structure schematic diagram, Fig. 2 is the principle schematic of the cell capture device capture cell of one embodiment of the invention.
As shown in Fig. 1 (A), Fig. 1 (B) and Fig. 2, a kind of large scale network array unicellular capture micro-fluidic device, including
Cell capture cavity 101.Microscope slide 102 it is provided with on the downside of cell capture cavity.It is provided with inflow on cell capture cavity 101
Mouth 103 and flow export 104.Cell capture device 105 it is provided with in cell capture cavity 101.Cell capture device 105 includes many
Individual unicellular capturing unit, multiple unicellular capturing units are connected with each other composition preset shape according to preset structure.Each slender
Born of the same parents' capturing unit includes capturing runner 203 and bypass runner 202.Be provided with slot on capture runner 203, slot with to be captured carefully
The size of born of the same parents matches.Capture runner 203 and the bypass runner 202 of each unicellular capturing unit are single with adjacent unicellular capture
The capture runner 203 of unit all connects with bypass runner 202.
In one embodiment of the invention, unicellular capturing unit includes a capture runner 203 and two by-passes
Road 202, two bypass runners constitute a rectangular configuration, and capture runner 203 connects one group of diagonal angle of rectangular configuration.Ordinary circumstance
Under, square structure has bigger facility than rectangular configuration.Particularly after capture runner 203 captures cell, owing to using
Square structure enables to two bypass runners 202 and shares the negative pressure of relative equilibrium so that unicellular capturing unit is again
When running into cell, flow to other adjacent unicellular capturing unit by bypass runner 202 equilibrium.
In one embodiment of the invention, cell capture device is matrix structure, and multiple unicellular capturing units are respectively
It is arranged on the different element positions of matrix structure.On each element position of matrix structure, corresponding setting unicellular
Capturing unit can be arranged as required to into or without, thus allow the different shape of multiple captured unicellular composition, in order to
In media environment, continued growth forms different structures.
In one embodiment of the invention, when unicellular capturing unit does not captures cell, the volume of capture runner 203
Flow rate is more than the volume flow rate of bypass runner 202, it is ensured that when capturing runner 203 and not capturing cell, have relative to side
The negative-pressure adsorption power that road runner 202 is bigger, so that capture runner 203 is easier to capture cell.
In one embodiment of the invention, being provided with negative pressure and provide device at flow export, negative pressure provides device to be used for
Between inflow entrance 103, cell capture device 105 and flow export 104, form negative pressure, make cell suspending liquid evenly into each
Unicellular capturing unit provides negative-pressure adsorption power.
In one embodiment of the invention, negative pressure provides device to include polyethylene tube 106 and syringe 108, by taking out
Syringe inflow entrance, between cell capture device and flow export formed negative pressure.Carried by polyethylene tube 106 and syringe 108
Simple, the low cost for negative pressure structure.
In one embodiment of the invention, cell capture cavity 101 is by being born photoetching rubber moulding by the SU8 of photoetching making
Take off film after uniformly coating a strata dimethyl siloxane drying on plate to make.
For making it is further understood that the present invention, will be described in detail by following example.
As shown in Fig. 1 (A) and Fig. 1 (B), the large scale network array unicellular capture micro-fluidic device of the embodiment of the present invention
Including cell capture cavity 101, microscope slide 102, inflow entrance 103, flow export 104, cell capture device 105, polyethylene tube
106, cell suspending liquid injection device 107 and syringe 108.With liquid-transfering gun, the cell suspending liquid after suitably dilution is instilled inflow
Mouth 103, adds negative pressure drainage by the syringe 108 pulling outlet to be connected to liquid, and cell is thin along with cell suspending liquid enters
In born of the same parents' acquisition equipment 105.
As in figure 2 it is shown, cell suspending liquid flows to from entrance 201, capture grid outlet 204 outflow, each unicellular capture
Unit is formed with two bypass runners 202 by capturing runner 203, is internally provided with slot at capture runner, can limit cell stream
Go out this unit.Each unicellular capturing unit, before capture cell, captures volume flow rate Q1 of runner, the volume flow of bypass runner
Rate is respectively Q2, Q3, captures volume flow rate Q1 of runner after capture ', the volume flow rate of bypass runner is respectively Q2 ', Q3 ',
Define each unit catch unicellular before volume flow rate than rq=Q1/max (Q2, Q3), catch unicellular after volume flow rate
Than rq '=Q1 '/max (Q2 ', Q3 ').
Fig. 3 is the schematic diagram of the theoretical model structure of one embodiment of the invention.As shown in Figure 3, it is illustrated that unicellular capture
Unit is arbitrary unit chosen in cell capture device.Along the direction of drop of pressure, A is the starting point of this unit, and B is this list
The terminal of unit, has three less paths between A to B and may select, respectively path 1, path 2, path 3.Have on path 1
There is the slot catching cell, and path 2,3 is only used as bypassing runner.The volume flow rate in path 1 is Q1.First segment body in path 2
Long-pending flow rate is Q21, second segment volume flow rate is Q22, Q21And Q22The most identical, its ratioCan be along with cell position
Cell capture state in distribution and runner changes.The first paragraph volume flow rate in path 3 is Q31, second segment volume flow rate is
Q32, Q31And Q32The most identical, its ratioCan be along with the cell capture state in cell position distribution and runner
Change.In path 1, a length of L of A, B point-to-point transmission1, the width of wider runner section is W1, for capturing the stream that cell is narrower
The a length of L of road partT, width is W10.In path 2, a length of L of first paragraph21, width of flow path is W2, second segment runner is long
Degree is L22, width is W2.In path 3, a length of L of first paragraph31, width of flow path is W3, second segment flow channel length is L32, width
For W3.After cell enters this unit, the runner that meeting prioritizing selection volume flow rate is bigger, for making cell preferentially enter capture runner
Select path 1, should makeCarry out theoretical derivation according to laminar model, can obtain the most succinct
Theory Solution, i.e. rQ=rQ1(1+min(λ2,λ3)) > 1, rQBy rQ1,λ2,λ3Determine, according to following expression formula,
rQ1Only determined by the geometric parameter of runner, as long as ensureing that the design of runner meets rQ1> 1 condition can ensure nothing
Opinion λ2,λ3The state change how to capture with peripheral unit and runner inner cell, all can meet's
Contact conditions.I.e. represent cell and can capture runner by prioritizing selection, after captured, Q1It is decreased to close to 0,No longer be satisfied, then subsequent cell can enter bypass runner, it is ensured that in capture runner in theory
Only capture unicellular.
Fig. 4 is the unicellular capture micro-fluidic device acquisition procedure schematic diagram of the present invention.As shown in Figure 4, along with cell with
Cell suspending liquid sequentially enters in runner, and cell suspending liquid flows into from the entrance of cell capture device, flows through cell capture device,
Cell selects the runner with minimum flow resistance by adaptivity, and is trapped in each unicellular capturing unit specific location,
Until after the capture of all unicellular capturing units is unicellular, unwanted cell flows out from end outlet again.Fig. 5 is one reality of the present invention
Execute 5 × 5 a kind of possible cell capture sequential schematic of cell capture device of example.As it is shown in figure 5, multiple cells are caught at cell
Obtain device to be captured by corresponding unicellular capturing unit successively.
Fig. 6 (A) is the capturing unit network simulation analysis chart of special cells array pattern under laminar flow condition.Fig. 6 (B) is layer
The capturing unit network simulation analysis chart of special cells array pattern under the conditions of stream.By filling capture runner 203 changing section
Cellular construction, then can change unicellular array arrangement mode, forms special cells array pattern, by the change of simulating, verifying structure
Change and still ensure that cell capture efficiency.Fig. 6 (C) is Fig. 6 (A) experimental result picture, basically identical with designed structure, but due to carefully
The phenomenons such as blocking that born of the same parents group's (note be not individual cells) causes and cell adhesion, cause subregion to exist with designed image
Different.Fig. 6 (D) is Fig. 6 (B) experimental result picture, basically identical with designed structure, but small part unit exists capture not
To single celled probability, there is some difference with designed image to cause subregion, with in A design compared with areas of blockage and
The situations such as cell adhesion reduce.
The large scale network array unicellular capture micro-fluidic device of the embodiment of the present invention, after capture, cellular array distribution can
Realize Small Distance, target cell also can guarantee that high capture rate in the case of original solution content is few, simultaneously effective slow down by
The device component failure that causes of blocking and can not reusable situation.
It addition, the large scale array of the embodiment of the present invention unicellular capture micro-fluidic device other constitute and act on right
It is all known for those skilled in the art, in order to reduce redundancy, does not repeats.
In the description of this specification, reference term " embodiment ", " some embodiments ", " example ", " specifically show
Example " or the description of " some examples " etc. means to combine this embodiment or example describes specific features, structure, material or spy
Point is contained at least one embodiment or the example of the present invention.In this manual, to the schematic representation of above-mentioned term not
Necessarily refer to identical embodiment or example.And, the specific features of description, structure, material or feature can be any
One or more embodiments or example in combine in an appropriate manner.
Although an embodiment of the present invention has been shown and described, it will be understood by those skilled in the art that: not
These embodiments can be carried out multiple change in the case of departing from the principle of the present invention and objective, revise, replace and modification, this
The scope of invention is limited by claim and equivalent thereof.
Claims (6)
1. a large scale network array unicellular capture micro-fluidic device, it is characterised in that include cell capture cavity, described
It is provided with inflow entrance and flow export on cell capture cavity, in described cell capture cavity, is provided with cell capture device, described
Cell capture device includes that multiple unicellular capturing unit, the plurality of unicellular capturing unit are connected with each other according to preset structure
Composition presets network shape, and each described unicellular capturing unit includes capturing runner and bypass runner, on described capture runner
Being provided with slot, described slot matches with the size of cell to be captured, the capture runner of each described unicellular capturing unit
All connect with capture runner and the bypass runner of adjacent unicellular capturing unit with bypass runner.
Large scale network array the most according to claim 1 unicellular capture micro-fluidic device, it is characterised in that described list
Cell capture unit includes that a described capture runner and two described bypass runners, two described bypass runners constitute a square
Shape structure, one group of diagonal angle of rectangular configuration described in described capture flow passage.
Large scale network array the most according to claim 2 unicellular capture micro-fluidic device, it is characterised in that described carefully
Born of the same parents' acquisition equipment is matrix structure, and multiple described unicellular capturing units are separately positioned on the different array element positions of described matrix structure
Put.
4., according to the large scale network array unicellular capture micro-fluidic device described in any one of claim 1-3, its feature exists
In, when described unicellular capturing unit does not captures cell, the volume flow rate of described capture runner is more than described bypass runner
Volume flow rate.
5., according to the large scale network array unicellular capture micro-fluidic device described in any one of claim 1-3, its feature exists
In, at described flow export, it being provided with negative pressure device is provided, described negative pressure provides device at described inflow entrance, described cell
Negative pressure is formed between acquisition equipment and described flow export.
Large scale network array the most according to claim 5 unicellular capture micro-fluidic device, it is characterised in that described negative
Pressure provides device to include polyethylene tube and syringe, is filled at described inflow entrance, described cell capture by syringe described in pull
Put formation negative pressure between described flow export.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107338183A (en) * | 2017-06-30 | 2017-11-10 | 北京酷搏科技有限公司 | Cell capture device |
| CN109576153A (en) * | 2018-11-09 | 2019-04-05 | 清华大学 | The method of cell electro' asion culture chip and device, cell electro' asion |
| WO2020093374A1 (en) * | 2018-11-09 | 2020-05-14 | 清华大学 | Cell electrofusion culture chip and device, and cell electrofusion method |
| WO2021056651A1 (en) * | 2019-09-25 | 2021-04-01 | 中国科学院苏州生物医学工程技术研究所 | Ultra-high-flux single-cell nucleic acid molecule real-time fluorescence quantitative analysis method, chip and system |
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| CN105219642A (en) * | 2015-09-09 | 2016-01-06 | 清华大学 | Catch and the micro-fluidic device of three-dimensional rotation for unicellular |
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| WO2011129651A3 (en) * | 2010-04-15 | 2012-04-05 | 주식회사 싸이토젠 | Microfluidic device and method for isolating target using same |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107338183A (en) * | 2017-06-30 | 2017-11-10 | 北京酷搏科技有限公司 | Cell capture device |
| CN109576153A (en) * | 2018-11-09 | 2019-04-05 | 清华大学 | The method of cell electro' asion culture chip and device, cell electro' asion |
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| CN106281965B (en) | 2019-05-31 |
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