CN206512179U - A kind of micro-fluidic chip that cell is sorted for laser - Google Patents
A kind of micro-fluidic chip that cell is sorted for laser Download PDFInfo
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- CN206512179U CN206512179U CN201720041038.2U CN201720041038U CN206512179U CN 206512179 U CN206512179 U CN 206512179U CN 201720041038 U CN201720041038 U CN 201720041038U CN 206512179 U CN206512179 U CN 206512179U
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- 238000000926 separation method Methods 0.000 claims abstract description 31
- 239000013307 optical fiber Substances 0.000 claims abstract description 19
- 210000004027 cell Anatomy 0.000 description 96
- 238000000034 method Methods 0.000 description 7
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- 238000004720 dielectrophoresis Methods 0.000 description 5
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- 238000012864 cross contamination Methods 0.000 description 1
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Abstract
The utility model is a kind of micro-fluidic chip that cell is sorted for laser.The chip enters liquid pool, buffer solution passage, cell solution including buffer solution, and to enter liquid pool, microelectrode, cell solution passage, electrode channel, the first top waste fluid channel, waste liquid pool on first, main channel, the second top waste fluid channel, waste liquid pool on second, the first cell separation passage, the first cell collecting pit, the second cell separation passage, the second cell collecting pit, second time waste liquid pool, the second bottom waste fluid channel, optical-fibre channel, first time waste liquid pool and the first bottom waste liquid logical.Described electrode channel is internally provided with microelectrode, and microelectrode is made up of two trapezoidal electrodes, is distributed up and down.The utility model requires very low to the feed speed of discrete phase, and the unicellular sequence speed of generation is more controllable, is conducive to post laser to the driven in registry of cell.
Description
Technical field
The utility model is related to biomedical technology, specifically generates unicellular sequence using dielectrophoresis, recycles laser
Trapping stiffness change sideways against cell cell movement locus micro-fluidic chip.
Background technology
Microflow control technique is the accurate control of the one kind risen 1980s and observing and controlling minute yardstick fluid, especially refers in particular to Asia
The technology of micrometer structure.Technique has sample volume small, and processing time is short, the advantages of frequency multiplexing technique and high surface to volume ratio.Especially
It is that it reacts fast, and sample consumption is few, microflow control technique is had well in biomedical aspect the characteristics of no cross contamination
Development prospect.
It is reported that the incidence of disease of cancer is all increasing year by year always, this causes us the research to circulating tumor cell to get over
More to pay close attention to.The research method of circulating tumor cell based on microflow control technique is also to emerge in an endless stream, to circulating tumor cell
Sorting is even more the most important thing, and comprehensive sorting both at home and abroad to circulating tumor cell at present, is uniquely supervised by U.S.'s food and medicine
The circulating tumor cell separation and number system for superintending and directing management board (FDA) certification are Cell Search, and its efficiency is 80%,
Nagrath etc. is devised captures chip with mini column array structure CTCs, has tested different cancerous cell line Ep content-addressable memorys up to level,
PBS suspension capture rates for Non-small cell lung carcinoma cell line NCI-H165 are up to 99%, Lv etc. and devise a kind of spacing
Gradual change type captures chip, and the effect that the chip structure is divided into two part filtration fractions of filtering and capture is filtered in blood
Impurity, reduces chip and blocks to greatest extent；Capture portion then devises different spacing microtrabeculaes, and spacing is successively decreased from 12 μm to 4 μm, blood
Cell can be by microtrabeculae, and tumour cell is then captured.Zhao Enming etc. is had been realized in granules of polystyrene and yeast cells
Capture and the effect such as ejection, and probed into mechanical function of the trapping stiffness to different size particles.
Utility model content
Not enough according to prior art, the technical problem that the utility model is intended to solve is to provide one kind using dielectrophoresis generation
Unicellular sequence, the micro-fluidic chip for recycling the trapping stiffness driving cell movement track of laser to deflect.Current chip
The feed speed of sheath stream, control discrete phase and continuous phase is typically all generated using cross passage and Y-shaped passage, so that raw
Into unicellular sequence.This chip adds a microelectrode being made up of two trapezoidal electrodes in main channel front portion, when cell is molten
Liquid flows through microelectrode, and the alternating current for being added to microelectrode two ends produces dielectrophoretic force, when positive dielectrophoresis effect occurs for cell, gradually
Move, be finally stuck at electrode edge to electrode edge, arranged along electrode edge, so that generating unicellular sequence is used for subsequently
Laser Driven.For generation sheath stream, the sample introduction of unicellular sequence pair continuous phase and discrete phase is generated using dielectrophoresis
The requirement of speed is very low, and the speed of unicellular sequence in the channel is controlled very well, is easy to post laser to drive cell more preferably to carry out；
The requirement to laser power can be reduced by introducing embedded optical fiber.Using compatible preferably multimode fibre, and according to emulation with
Actual conditions have carried out structure optimization to passage, complete the design and making of overall chip, are easy to operation.The chip has height
Flux, it is unmarked pollution-free, it is adaptable to the features such as practical application.
The technical solution of the utility model is：
It is a kind of for laser sort cell micro-fluidic chip, the chip include buffer solution enter liquid pool, buffer solution passage, carefully
Cell lysis liquid enters liquid pool, microelectrode, cell solution passage, electrode channel, the first top waste fluid channel, waste liquid pool on first, main logical
Road, the second top waste fluid channel, waste liquid pool, the first cell separation passage, the first cell collecting pit, the second cell separation on second
Passage, the second cell collecting pit, second time waste liquid pool, the second bottom waste fluid channel, optical-fibre channel, first time waste liquid pool and first
Bottom waste liquid leads to.
Wherein, cell solution passage, electrode channel are connected with main channel successively level；It is logical in cell solution passage and electrode
The junction in road, is also associated with buffer solution passage, and buffer solution passage has two passes, respectively positioned at the top of cell solution passage
And bottom, and equal and cell solution passage angle at 45 °；Electrode channel and the junction of main channel, have been also respectively connected with first
Top waste fluid channel and the first bottom waste liquid are logical, and the two is located at the upper and lower part of main channel respectively, and equal and main channel is at 45 °
Angle；The end of main channel, from top to bottom, is connected with the second top waste fluid channel in turn, the first cell separation passage, second thin
Born of the same parents' split tunnel and the second bottom waste fluid channel, wherein, the angle of the adjacency channel of four branched bottoms is 30 °, and second
Cell separation passage and main channel level connection joint；80 μm of bottom, is also associated with optical-fibre channel before the end of main channel.
The two passes of described buffer solution passage are intersected in buffer solution and enter liquid pool, and cell solution feeder connection is connected to cell
Solution enters liquid pool.
The end of the first described top waste fluid channel is connected to waste liquid pool on first；The end of second top waste fluid channel connects
There is waste liquid pool on second；The end of first cell separation passage is connected to the first cell collecting pit；The end of second cell separation passage
It is terminated with the second cell collecting pit；The end of second bottom waste fluid channel is connected to second time waste liquid pool；What the first bottom waste liquid led to
End is connected to first time waste liquid pool.
Described electrode channel is internally provided with microelectrode, and microelectrode is made up of two trapezoidal electrodes, is distributed, passes through up and down
Respective vertical electrode is respectively connected to the two ends of AC power；The long 1.2mm of whole microelectrode, wide 180 μm, two trapezoidal electrodes are left
It is 80 μm to hold spacing, and right-hand member spacing is 40 μm.
The width of electrode channel is 240 μm, and other channel widths are 100 μm.
The beneficial effects of the utility model are：
The utility model generates unicellular sequence with dielectrophoresis, and unicellular sequence is generated relative to traditional sheath stream,
This chip requires very low to the feed speed of discrete phase, and the unicellular sequence speed of generation is more controllable, is conducive to post laser
To the driven in registry of cell.Chip structure is ingenious in design, and feature is strong, can not only realize cell sorting, can also be effective
Cell count is realized, without any mark, unmarked, high-throughout cell sorting is realized.
Brief description of the drawings
Fig. 1 illustrates for the utility model chip structure.
Fig. 2 is chip electrode passage enlarged diagram.
Wherein, 1：Buffer solution enters liquid pool；2：Buffer solution passage；3：Cell solution enters liquid pool；4：Microelectrode；5：Cell solution
Passage；6：Electrode channel；7：First top waste fluid channel；8：Waste liquid pool on first；9：Main channel；10：Second top waste liquid leads to
Road；11：Waste liquid pool on second；12：First cell separation passage；13：First cell collecting pit；14：Second cell separation passage；
15：Second cell collecting pit 16：Second time waste liquid pool；17：Second bottom waste fluid channel；18：Optical-fibre channel 19：First time waste liquid
Pond；20：First bottom waste fluid channel.
Embodiment
The utility model is further described with reference to embodiment and its accompanying drawing.
The structure of the micro-fluidic chip described in the utility model that cell is sorted for laser is as shown in figure 1, including buffering
Liquid enter liquid pool 1, buffer solution passage 2, cell solution enter liquid pool 3, microelectrode 4, cell solution passage 5, on electrode channel 6, first
Waste liquid pool 8 in portion's waste fluid channel 7, first, main channel 9, waste liquid pool 11, the first cell in the second top waste fluid channel 10, second
Split tunnel 12, the first cell collecting pit 13, the second cell separation passage 14, the second cell collecting pit 15, second time waste liquid pool
16th, the second bottom waste fluid channel 17, optical-fibre channel 18, first time bottom waste fluid channel 20 of waste liquid pool 19 and first.Whole chip
All passages, the thickness for entering liquid pool, collecting pit and waste liquid pool are all equal.
Wherein, cell solution passage 5, electrode channel 6 are connected with main channel 9 successively level；In cell solution passage 5 and electricity
The junction of pole passage 6, is also associated with buffer solution passage 2, and buffer solution passage 2 has two passes, logical positioned at cell solution respectively
The upper and lower part in road 5, and the equal and angle at 45 ° of cell solution passage 5；Electrode channel 6 and the junction of main channel 9, also divide
The first top waste fluid channel 7 and the first bottom waste liquid logical 20 are not connected with, and the two is located at the upper and lower part of main channel 9 respectively,
And the equal and angle at 45 ° of main channel 9；The end of main channel 9, from top to bottom, is connected with the second top waste fluid channel 10, in turn
One cell separation passage 12, the second cell separation passage 14 and the second bottom waste fluid channel 17, wherein, the phase of four branched bottoms
The angle of adjacent passage is 30 °, and the second cell separation passage 14 and the level connection joint of main channel 9；80 μ before the end of main channel 9
M bottom, is also associated with optical-fibre channel 18.
The two passes of described buffer solution passage 2 are intersected in buffer solution and enter liquid pool 1, and the entrance of cell solution passage 5 is connected to
Cell solution enters liquid pool 3.
The end of the first described top waste fluid channel 7 is connected to waste liquid pool 8 on first；The end of second top waste fluid channel 10
It is terminated with waste liquid pool 11 on second；The end of first cell separation passage 12 is connected to the first cell collecting pit 13；Second cell point
End from passage 14 is connected to the second cell collecting pit 15；The end of second bottom waste fluid channel 17 is connected to second time waste liquid pool
16；The end of first bottom waste liquid logical 20 is connected to first time waste liquid pool 19.
Described electrode channel 6 is internally provided with microelectrode 4, and microelectrode 4 is made up of two trapezoidal electrodes, is distributed up and down,
The two ends of AC power are respectively connected to by respective vertical electrode；The long 1.2mm of whole microelectrode, wide 180 μm, two trapezoidal electricity
Ultra-Left end (arrival end) spacing is 80 μm, and right-hand member spacing (port of export) is that " a ladder is formed between 40 μm, such two electrode
Shape " passage.
The width of electrode channel 6 is 240 μm, and other channel widths are 100 μm.
Described buffer solution passage 2 has two passes, respectively positioned at the upper and lower part of cell solution passage 5, and and
The angle at 45 ° of cell solution passage 5, buffer solution passage 2 mainly serves for ensuring cell solution and flows through two microelectrodes 4.Electrode leads to
Road 6 and the junction of main channel 9, have been also respectively connected with the first top waste fluid channel 7 and the first bottom waste liquid logical 20, the two point
Not Wei Yu main channel 9 upper and lower part, and and the angle at 45 ° of main channel 9, the first top waste fluid channel 7 and the first bottom
Waste fluid channel 20 can discharge the buffer solution waste liquid in electrode channel 9, and fiber perpendicular insertion is apart from 80 μm of main channel end
It is the flowing of unicellular sequence in position, main channel 9, when cell passes through optical-fibre channel 18, the laser in optical fiber is to cell one
Fixed trapping stiffness, makes cell deflect, and the waste liquid in main channel 9 is logical from the second top waste fluid channel 10 and the second bottom waste liquid
Road 17 is flowed out, and the first cell separation passage 12 and the second cell separation passage 14 are flowed to respectively by the cell of different deflections, from
And realize the sorting of cell.
Microelectrode 4 is made up of the trapezoidal electrode of about two distributions, and alternating current is respectively connected to by respective vertical electrode
The two ends in source；The end of first top waste fluid channel is that waste liquid pool on first, the first bottom waste fluid channel end are to give up for first time
Liquid pool, the end of the second top waste fluid channel be waste liquid pool on second, the second bottom waste fluid channel end be second time waste liquid pool,
The end of first cell separation passage is that the first cell collecting pit, the end of the second cell separation passage are that the second cell is collected
Pond.
Embodiment 1
In the utility model, described buffer solution passage 2 be divided into above and below two passes be connected respectively with cell solution passage 5
And the angle with cell solution passage 5 is 45 °, the two passes of buffer solution passage 2 are intersected in buffer solution and enter liquid pool 1, by delaying
Fliud flushing enters liquid pool 1 and enters buffer solution；The entrance connection cell solution of cell solution passage enters liquid pool 3, and buffer solution passage 2 and cell are molten
The end of the intersection of liquid passage 5 is connection electrode passage 6, the long 2mm of electrode channel 6, and wide 240 μm, microelectrode 4 is trapezoidal by two
Electrode composition (is ITO electrode.), the whole long 1.2mm of microelectrode is wide 180 μm, between two trapezoidal electrode left ends (arrival end)
Away from for 80 μm, right-hand member spacing (port of export) is to form " trapezoidal " passage between 40 μm, two electrodes, by two electrodes each
Vertical electrode be respectively connected to the two ends of AC power.Between electrode channel 6 and main channel 9 by the first top waste fluid channel 7 and
First bottom waste fluid channel 20 is connected, i.e., cell solution passage 5, electrode channel 6 are connected with main channel 9 successively level, main channel
The angle of long 16mm, the first top waste fluid channel 7 and the first bottom waste fluid channel 20 and main channel 9 is 45 °, and cell passes through this
The intersection of two waste fluid channels and main channel enters main channel 9, and main channel 9 is unicellular sequence, and the width of main channel 9 is
100μm；The end of main channel 9 separates the branched bottom to form four aplysia punctata connections, and the second top is followed successively by from top to bottom and is given up
Liquid passage 10, the first cell separation passage 12, the second cell separation passage 14 and the second bottom waste fluid channel 17, such as chip structure
Shown in figure, the second cell separation passage 14 connects with the level of main channel 9, and four branched bottom mutual angles are 30 °, Ta Menkuan
Degree is equal, is 100 μm, channel thickness is also 100 μm.What four branched bottom ends were sequentially connected from the top down is second
Upper waste liquid pool 11, the first cell collecting pit 13, the second cell collecting pit 15 and second time waste liquid pool 16.Using poly- in this experiment
Dimethyl siloxane (PDMS) as chip material, i.e. buffer solution enter liquid pool 1, buffer solution passage 2, cell solution enter liquid pool 3, it is thin
Born of the same parents' solution channel 5, electrode channel 6, waste liquid pool 8, main channel 9, the first bottom waste liquid lead in the first top waste fluid channel 7, first
Road 20, second time waste liquid pool 19, waste liquid pool 11, the first cell separation passage 12, first in the second top waste fluid channel 10, second
Cell collecting pit 13, the second cell separation passage 14, the second cell collecting pit 15, the second bottom waste fluid channel 17, second time give up
Liquid pool 16, optical-fibre channel 18 are PDMS.Microelectrode 4 uses ITO electrode.In the chips reserve optical-fibre channel 18, optical fiber it is straight
Footpath be 125 μm, it is desirable to fiber end face perpendicular alignmnet main channel 9, then optical fiber is embedded into optical fiber reserved passageway, vacuumize by
Resin glue is drawn into optical fiber reserved passageway and sealed.
The operation principle and process of the utility model chip be：Cell solution passage sample introduction into electrode channel, buffer solution
Passage is used for ensureing that cell solution all flows through electrode, two electrodes is connected with AC power two ends respectively, so in electrode
Between can produce dielectrophoretic force, use the yeast cell solution that electrical conductivity is 60uS/cm, alternating current voltage 8V or so, frequency
50MHz or so, now occurs positive dielectrophoretic force, and cell is close to electrode edge.Unicellular sequence is formed, electrode channel both sides
Waste liquid is flowed out by the first top waste fluid channel and the first bottom waste fluid channel respectively, and cellular sequences enter main channel, along streamline
Direction enter the position that optical fiber is embedded in and irradiated by the laser in optical fiber because different cell different cell membrane and cell
Matter, so power suffered under being irradiated for the laser of different wave length, power is also different, therefore this chip can be tested to difference
The cell of species and size is sorted, and the cell by sorting, which eventually enters into cell separation passage, the collection of cell collecting pit,
Complete sorting.
The utility model does not address part and is applied to prior art.
Claims (2)
1. a kind of micro-fluidic chip that cell is sorted for laser, it is characterized in that the chip enters liquid pool, buffer solution including buffer solution
Passage, cell solution enter liquid pool, microelectrode, cell solution passage, electrode channel, the first top waste fluid channel, waste liquid on first
Pond, main channel, the second top waste fluid channel, waste liquid pool, the first cell separation passage, the first cell collecting pit, second on second
Cell separation passage, the second cell collecting pit, second time waste liquid pool, the second bottom waste fluid channel, optical-fibre channel, first time waste liquid
Pond and the first bottom waste liquid are logical；
Wherein, cell solution passage, electrode channel are connected with main channel successively level；In cell solution passage and electrode channel
Junction, is also associated with buffer solution passage, and buffer solution passage has two passes, respectively positioned at the top of cell solution passage with
Portion, and equal and cell solution passage angle at 45 °；Electrode channel and the junction of main channel, have been also respectively connected with the first top
Waste fluid channel and the first bottom waste liquid are logical, and the two is located at the upper and lower part of main channel, and equal and main channel folder at 45 ° respectively
Angle；The end of main channel, from top to bottom, is connected with the second top waste fluid channel, the first cell separation passage, the second cell in turn
Split tunnel and the second bottom waste fluid channel, wherein, the angle of the adjacency channel of four branched bottoms is 30 °, and second is thin
Born of the same parents' split tunnel and main channel level connection joint；80 μm of bottom, is also associated with optical-fibre channel before the end of main channel；
The two passes of described buffer solution passage are intersected in buffer solution and enter liquid pool, and cell solution feeder connection is connected to cell solution
Enter liquid pool；
The end of the first described top waste fluid channel is connected to waste liquid pool on first；The end of second top waste fluid channel is connected to
Waste liquid pool on two；The end of first cell separation passage is connected to the first cell collecting pit；The end of second cell separation passage connects
There is the second cell collecting pit；The end of second bottom waste fluid channel is connected to second time waste liquid pool；The logical end of first bottom waste liquid
It is connected to first time waste liquid pool；
Described electrode channel is internally provided with microelectrode, and microelectrode is made up of two trapezoidal electrodes, is distributed up and down, by respective
Vertical electrode be respectively connected to the two ends of AC power；The long 1.2mm of whole microelectrode, wide 180 μm, two trapezoidal electrode left ends
Spacing is 80 μm, and right-hand member spacing is 40 μm.
2. the micro-fluidic chip of cell is sorted for laser as claimed in claim 1, it is characterized in that the width of electrode channel is
240 μm, other channel widths are 100 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720041038.2U CN206512179U (en) | 2017-01-13 | 2017-01-13 | A kind of micro-fluidic chip that cell is sorted for laser |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720041038.2U CN206512179U (en) | 2017-01-13 | 2017-01-13 | A kind of micro-fluidic chip that cell is sorted for laser |
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| CN206512179U true CN206512179U (en) | 2017-09-22 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735429A (en) * | 2019-01-28 | 2019-05-10 | 佛山市铬维科技有限公司 | Micro-fluidic chip and the system and its separation method for separating various kinds of cell |
| CN111644212A (en) * | 2020-05-22 | 2020-09-11 | 华东理工大学 | Micro-fluidic chip and nano-particle separation device |
-
2017
- 2017-01-13 CN CN201720041038.2U patent/CN206512179U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735429A (en) * | 2019-01-28 | 2019-05-10 | 佛山市铬维科技有限公司 | Micro-fluidic chip and the system and its separation method for separating various kinds of cell |
| CN111644212A (en) * | 2020-05-22 | 2020-09-11 | 华东理工大学 | Micro-fluidic chip and nano-particle separation device |
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