CN106281965B - The unicellular capture micro-fluidic device of large scale network array - Google Patents
The unicellular capture micro-fluidic device of large scale network array Download PDFInfo
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- CN106281965B CN106281965B CN201610671600.XA CN201610671600A CN106281965B CN 106281965 B CN106281965 B CN 106281965B CN 201610671600 A CN201610671600 A CN 201610671600A CN 106281965 B CN106281965 B CN 106281965B
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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 unicellular capture micro-fluidic devices of large scale network array, including cell capture cavity, inflow entrance and outflux are provided on the cell capture cavity, cell capture device is provided in the cell capture cavity, the cell capture device includes multiple unicellular capturing units, the multiple unicellular capturing unit is connected with each other composition preset shape according to preset structure, each unicellular capturing unit includes capture runner and bypass runner, slot is provided on the capture runner, the size of the slot and cell to be captured matches, the capture runner and bypass runner of each unicellular capturing unit are connected to the capture runner of adjacent unicellular capturing unit and bypass runner.The present invention has the advantage that network-like distribution is presented in unicellular capturing unit, multi-pass line structure when capture is realized, and there is adaptivity, capture rate can be improved, while reducing the risk of device blocking.
Description
Technical field
The present invention relates to cell capture technical fields, and it is micro-fluidic specifically to design a kind of unicellular capture of large scale network array
Device.
Background technique
Currently, unicellular capture device in the related technology passes through a large amount of capturing unit of arrangement in runner, or make
With the mode of multi-channel parallel, to improve flux.After cell suspending liquid injection, cell being caught into what is designed in a random way
Unit is obtained, a small amount of cell is limited in runner by the groove in slot or runner, and other most cells are with suspension flow
It exports.But a kind of Gaussian Profile is presented in the probability that can cell enter capturing unit in runner, therefore reduces such
The capturing efficiency of mode needs high-throughput cell to flow into the cell capture rate that could guarantee each unit, in addition most devices
Using the serial connection mode of capturing unit, device fault, the risk that cannot be reused caused by causing because of blocking.
Summary of the invention
The present invention is directed at least solve one of above-mentioned technical problem.
For this purpose, being realized it is an object of the invention to propose a kind of unicellular capture micro-fluidic device of large scale network array
Multi-pass line structure when capture, and there is adaptivity, capture rate can be improved, while reducing the risk of device blocking.
To achieve the goals above, embodiment of the invention discloses a kind of unicellular capture miniflows of large scale network array
Device, including cell capture cavity are controlled, is provided with inflow entrance and outflux, the cell capture chamber on the cell capture cavity
It is provided with cell capture device in vivo, the cell capture device includes multiple unicellular capturing units, the multiple unicellular
Capturing unit according to preset structure be connected with each other composition preset shape, each unicellular capturing unit include capture runner and
Runner is bypassed, is provided with slot on the capture runner, the size of the slot and cell to be captured matches, each list
The capture runner and bypass runner of cell capture unit connect with the capture runner of adjacent unicellular capturing unit and bypass runner
It is logical.
The unicellular capture micro-fluidic device of large scale network array according to an embodiment of the present invention, unicellular capturing unit are in
Multi-pass line structure when capture is realized in existing network-like distribution, and has adaptivity, capture rate can be improved, while reducing device
The risk of part blocking.
It, can be in addition, the unicellular capture micro-fluidic device of large scale network array according to the above embodiment of the present invention
With following additional technical characteristic:
Further, the unicellular capturing unit include a capture runner and two bypass runners, two
A bypass runner constitutes a rectangular configuration, and the capture runner be connected to the rectangular configuration one group is diagonal.
Further, the cell capture device is matrix structure, and multiple unicellular capturing units are separately positioned on
On the different element positions of the matrix structure.
Further, when the unicellular capturing unit does not capture cell, the volume flow rate of the capture runner is greater than
The volume flow rate of the bypass runner.
Further, it is provided with negative pressure at the outflux, device is provided, the negative pressure provides device and is used for described
Negative pressure is formed between inflow entrance, the cell capture device and the outflux.
Further, it includes polyethylene pipe and syringe that the negative pressure, which provides device, by syringe described in pull in institute
It states and forms negative pressure between inflow entrance, the cell capture device and the outflux.
Additional aspect and advantage of the invention will be set forth in part in the description, and will partially become from the following description
Obviously, or practice through the invention is recognized.
Detailed description of the invention
Above-mentioned and/or additional aspect of the invention and advantage will become from the description of the embodiment in conjunction with the following figures
Obviously and it is readily appreciated that, in which:
Fig. 1 (A) is the main view knot of the unicellular capture micro-fluidic device of the large scale network array of one embodiment of the invention
Structure schematic diagram；
Fig. 1 (B) is the side view knot of the unicellular capture micro-fluidic device of the large scale network array of one embodiment of the invention
Structure schematic diagram；
Fig. 2 is the schematic illustration of the cell capture device capture cell of one embodiment of the invention；
Fig. 3 is the schematic diagram of the theoretical model building 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 a kind of possible cell capture sequential schematic of 5 × 5 cell capture devices 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.
Specific embodiment
The embodiment of the present invention is described below in detail, examples of the embodiments are shown in the accompanying drawings, wherein from beginning to end
Same or similar label indicates same or similar element or element with the same or similar functions.Below with reference to attached
The embodiment of figure description is exemplary, and for explaining only the invention, and is not considered as limiting the invention.
In the description of the present invention, it is to be understood that, term " center ", " longitudinal direction ", " transverse direction ", "upper", "lower",
The orientation or positional relationship of the instructions such as "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outside" is
It is based on the orientation or positional relationship shown in the drawings, is merely for convenience of description of the present invention and simplification of the description, rather than instruction or dark
Show that signified device or element must have a particular orientation, be constructed and operated in a specific orientation, therefore should not be understood as pair
Limitation of the invention.In addition, term " first ", " second " are used for description purposes only, it is not understood to indicate or imply opposite
Importance.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected；It can
To be mechanical connection, it is also possible to be electrically connected；It can be directly connected, can also can be indirectly connected through an intermediary
Connection inside two elements.For the ordinary skill in the art, above-mentioned term can be understood at this with concrete condition
Concrete meaning in invention.
Referring to following description and drawings, it will be clear that these and other aspects of the embodiment of the present invention.In these descriptions
In attached drawing, some particular implementations in the embodiment of the present invention are specifically disclosed, to indicate to implement implementation of the 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, of the invention
Embodiment includes all changes, modification and the equivalent fallen within the scope of the spirit and intension of attached claims.
The unicellular capture micro-fluidic device of large scale network array according to an embodiment of the present invention is described below in conjunction with attached drawing.
Fig. 1 (A) is the main view knot of the unicellular capture micro-fluidic device of the large scale network array of one embodiment of the invention
Structure schematic diagram, Fig. 1 (B) are the side view knots of the unicellular capture micro-fluidic device of the large scale network array of one embodiment of the invention
Structure schematic diagram, Fig. 2 are the schematic illustrations 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 unicellular capture micro-fluidic device of large scale network array, including
Cell capture cavity 101.Glass slide 102 is provided on the downside of cell capture cavity.Inflow is provided on cell capture cavity 101
Mouth 103 and outflux 104.Cell capture device 105 is provided in cell capture cavity 101.Cell capture device 105 includes more
A unicellular capturing unit, multiple unicellular capturing units are connected with each other composition preset shape according to preset structure.It is each slender
Born of the same parents' capturing unit includes capture runner 203 and bypass runner 202.Slot is provided on capture runner 203, slot and to be captured thin
The size of born of the same parents matches.The capture runner 203 and bypass runner 202 of each unicellular capturing unit and adjacent unicellular capture are single
The capture runner 203 of member is connected to bypass runner 202.
In one embodiment of the invention, unicellular capturing unit includes a capture runner 203 and two by-pass
Road 202, two bypass runners constitute a rectangular configuration, and one group of the capture connection rectangular configuration of runner 203 is diagonal.Ordinary circumstance
Under, square structure has bigger convenience than rectangular configuration.Especially after capture runner 203 captures cell, due to using
Square structure enables to two to bypass the negative pressure that runner 202 shares relative equilibrium, so that unicellular capturing unit is again
When encountering cell, other adjacent unicellular capturing units are flowed to by bypassing 202 equilibrium of runner.
In one embodiment of the invention, cell capture device is matrix structure, multiple unicellular capturing unit difference
It is arranged on the different element positions of matrix structure.On each element position of matrix structure, corresponding setting it is unicellular
Capturing unit can according to need be set as with or without, to allow the different shape of multiple captured unicellular compositions, so as to
Continued growth forms different structures in media environment.
In one embodiment of the invention, when unicellular capturing unit does not capture cell, the volume of runner 203 is captured
Flow rate is greater than the volume flow rate of bypass runner 202, it is ensured that when capture runner 203 does not capture cell, has relative to side
The bigger negative-pressure adsorption power of road runner 202, to make to capture the more easily capture cell of runner 203.
In one embodiment of the invention, it is provided with negative pressure at outflux, device is provided, negative pressure provides device and is used for
Negative pressure is formed between inflow entrance 103, cell capture device 105 and outflux 104, makes cell suspending liquid evenly into each
Unicellular capturing unit provides negative-pressure adsorption power.
In one embodiment of the invention, it includes polyethylene pipe 106 and syringe 108 that negative pressure, which provides device, passes through pumping
Syringe forms negative pressure between inflow entrance, cell capture device and outflux.It is mentioned by polyethylene pipe 106 and syringe 108
It is simple for negative pressure structure, at low cost.
In one embodiment of the invention, cell capture cavity 101 passes through in the negative photoetching rubber moulding of SU8 by photoetching making
It takes off film after uniformly coating strata dimethyl siloxane drying on plate and is made.
To make it is further understood that the present invention, will be described in detail by following embodiment.
As shown in Fig. 1 (A) and Fig. 1 (B), the unicellular capture micro-fluidic device of the large scale network array of the embodiment of the present invention
Including cell capture cavity 101, glass slide 102, inflow entrance 103, outflux 104, cell capture device 105, polyethylene pipe
106, cell suspending liquid injection device 107 and syringe 108.The cell suspending liquid after appropriate dilution is instilled with liquid-transfering gun and is flowed into
Mouth 103 adds negative pressure drainage to liquid by the syringe 108 for pulling outlet connected, and cell is thin as cell suspending liquid enters
In born of the same parents' acquisition equipment 105.
As shown in Fig. 2, cell suspending liquid is flowed into from entrance 201, and 204 outflow of capture grid outlet, each unicellular capture
Unit is made of capture runner 203 and two bypass runners 202, is internally provided with slot in capture runner, can be limited cell stream
The unit out.Each unicellular capturing unit captures the volume flow rate Q1 of runner, bypasses the volume flow of runner before capturing cell
Rate is respectively Q2, Q3, and the volume flow rate Q1 ' of runner is captured after capture, and the volume flow rate for bypassing runner is respectively Q2 ', Q3 ',
It defines each unit and captures unicellular preceding volume flow rate ratio rq=Q1/max (Q2, Q3), the volume flow rate after capture is unicellular
Than rq '=Q1 '/max (Q2 ', Q3 ').
Fig. 3 is the schematic diagram of the theoretical model building of one embodiment of the invention.As shown in figure 3, illustrating unicellular capture
Unit is the unit of any selection in cell capture device.Along the direction of drop of pressure, A is the starting point of the unit, and B is the list
The terminal of member, three lesser paths are shared between A to B may be selected, respectively path 1, path 2, path 3.Have on path 1
There is the slot for capturing cell, and path 2,3 is only as bypass runner.The volume flow rate in path 1 is Q1.First segment body in path 2
Product flow rate is Q21, second segment volume flow rate is Q22, Q21And Q22It is not necessarily identical, ratioIt can be with cell position
Cell capture state in distribution and runner changes.The first segment volume flow rate in path 3 is Q31, second segment volume flow rate is
Q32, Q31And Q32It is not necessarily identical, ratioIt can be with the cell capture state in cell position distribution and runner
Change.In path 1, A, B point-to-point transmission length is L1, the width of wider runner section is W1, for capturing the relatively narrow stream of cell
The length of road part is LT, width W10.In path 2, the first segment length is L21, width of flow path W2, second segment runner is long
Degree is L22, width W2.In path 3, the first segment length is L31, width of flow path W3, second segment flow channel length is L32, width
For W3.After cell enters the unit, the runner that preferentially volume flow rate can be selected bigger, to make cell preferentially enter capture runner
Path 1 is selected, should be madeTheory deduction is carried out according to laminar model, it is available very succinct
Theory Solution, i.e. rQ=rQ1(1+min(λ2,λ3)) > 1, rQBy rQ1,λ2,λ3Determine, according to following expression formula it is found that
rQ1It is only determined by the geometric parameter of runner, as long as guaranteeing that the design of runner meets rQ1> 1 condition can guarantee nothing
By λ2,λ3The state change how to capture with peripheral unit and runner inner cell, can meet's
Contact conditions.Capture runner, after captured, Q can preferentially be selected by representing cell1It is decreased to close to 0,It is no longer satisfied, then subsequent cell can enter bypass runner, guarantee in capture runner theoretically
It only captures unicellular.
Fig. 4 is unicellular capture micro-fluidic device acquisition procedure schematic diagram of the invention.As shown in figure 4, with cell with
Cell suspending liquid sequentially enters in runner, and cell suspending liquid is flowed into from the entrance of cell capture device, flows through cell capture device,
Cell has the runner of minimum flow resistance by adaptivity selection, and is trapped in each unicellular capturing unit specific location,
Until unwanted cell is flowed out from end outlet again after all unicellular capturing unit captures are unicellular.Fig. 5 is a reality of the invention
Apply a kind of possible cell capture sequential schematic of 5 × 5 cell capture devices of example.As shown in figure 5, multiple cells are caught in cell
Device is obtained successively to be captured by corresponding unicellular capturing unit.
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.203 changing section of runner is captured by filling
Unicellular array arrangement mode then can be changed in cellular construction, forms special cells array pattern, passes through the change of simulating, verifying structure
Change still ensures that cell capture efficiency.Fig. 6 (C) is Fig. 6 (A) experimental result picture, almost the same with designed structure, but due to thin
Phenomena such as blocking caused by born of the same parents group's (attention is not individual cells) and cell adherence, partial region and designed image is caused to exist
Different.Fig. 6 (D) is Fig. 6 (B) experimental result picture, almost the same with designed structure, but small part unit has capture not
To single celled possibility, causing partial region with designed image, there is some difference, in A design compared with areas of blockage and
Situations such as cell adherence, is reduced.
The unicellular capture micro-fluidic device of the large scale network array of the embodiment of the present invention, cellular array distribution can after capture
Realize that small spacing, target cell also can guarantee high capture rate in the case where original solution content is few, simultaneously effective slow down by
Component failure caused by device blocks and the case where cannot reusing.
In addition, the other compositions and effect pair of the unicellular capture micro-fluidic device of the large scale array of the embodiment of the present invention
For those skilled in the art be all it is known, in order to reduce redundancy, do not repeat them here.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
Centainly refer to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be any
One or more embodiment or examples in can be combined in any suitable 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
A variety of change, modification, replacement and modification can be carried out to these embodiments in the case where being detached from the principle of the present invention and objective, this
The range of invention is by claim and its equivalent limits.
Claims (1)
1. a kind of unicellular capture micro-fluidic device of large scale network array, which is characterized in that described including cell capture cavity
It is provided with inflow entrance and outflux on cell capture cavity, cell capture device is provided in the cell capture cavity, it is described
Cell capture device includes multiple unicellular capturing units, and the multiple unicellular capturing unit is connected with each other according to preset structure
Default network shape is formed, each unicellular capturing unit includes capturing runner and bypass runner, on the capture runner
It is provided with slot, the size of the slot and cell to be captured matches, the capture runner of each unicellular capturing unit
With bypass runner and the capture runner of adjacent unicellular capturing unit and bypass runner and be connected to, wherein unicellular caught described
When obtaining unit and not capturing cell, the volume flow rate of the capture runner is greater than the volume flow rate of the bypass runner, in the stream
Exit is provided with negative pressure and provides device, the negative pressure provide device be used for the inflow entrance, the cell capture device and
Negative pressure is formed between the outflux, it includes polyethylene pipe and syringe that the negative pressure, which provides device, by injecting described in pull
Device forms negative pressure between the inflow entrance, the cell capture device and the outflux；The unicellular capturing unit packet
The capture runner and two bypass runners are included, two bypass runners constitute a rectangular configuration, described to catch
Obtain runner be connected to one group of the rectangular configuration it is diagonal, the cell capture device is matrix structure, multiple described unicellular to catch
Unit is obtained to be separately positioned on the different element positions of the matrix structure.
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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 |
| WO2020093374A1 (en) * | 2018-11-09 | 2020-05-14 | 清华大学 | Cell electrofusion culture chip and device, and cell electrofusion method |
| 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 |
| 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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| WO2011129651A3 (en) * | 2010-04-15 | 2012-04-05 | 주식회사 싸이토젠 | Microfluidic device and method for isolating target using same |
| CN104513787A (en) * | 2015-01-07 | 2015-04-15 | 东北大学 | Integrated micro-fluidic chip and system for capture, culture and administration of single cells |
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| WO2009130694A2 (en) * | 2008-04-21 | 2009-10-29 | Cell Kinetics Ltd. | Flat cell carriers with cell traps |
| CN105219642B (en) * | 2015-09-09 | 2018-02-09 | 清华大学 | For unicellular capture and the micro-fluidic device of three-dimensional rotation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2011129651A3 (en) * | 2010-04-15 | 2012-04-05 | 주식회사 싸이토젠 | Microfluidic device and method for isolating target using same |
| CN104513787A (en) * | 2015-01-07 | 2015-04-15 | 东北大学 | Integrated micro-fluidic chip and system for capture, culture and administration of single cells |
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