CN114713297B - Microfluidic Chip - Google Patents
Microfluidic Chip Download PDFInfo
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- CN114713297B CN114713297B CN202110003615.XA CN202110003615A CN114713297B CN 114713297 B CN114713297 B CN 114713297B CN 202110003615 A CN202110003615 A CN 202110003615A CN 114713297 B CN114713297 B CN 114713297B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
Abstract
A microfluidic chip includes a droplet inlet, a droplet outlet, a droplet incubation channel, a continuous phase channel, and a confluence channel. The droplet incubation channel is connected to the droplet inlet for the continuous phase and a plurality of droplets carried by the continuous phase to flow into the droplet incubation channel. The continuous communication channel is positioned on at least one side of the droplet incubation channel and is communicated with the droplet incubation channel through the opening, so that the continuous phase and the droplets enter the continuous phase channel through the opening after being separated, and the droplets are incubated in the droplet incubation channel. The continuous phase channel and the liquid drop incubation channel are intersected in the converging channel, so that the continuous phase and the incubated liquid drops are converged again at the converging channel. The liquid drop outlet is connected with the confluence channel. The application is beneficial to simplifying the chip structure and improving the universality while realizing the purposes of incubating and observing liquid drops.
Description
Technical Field
The application relates to the technical field of microfluidics, in particular to a microfluidic chip.
Background
In recent years, microfluidic systems have been widely used in the biological and chemical fields as a platform capable of integrating a variety of functional modules. Microfluidic chips, also known as lab-on-a-chip, can integrate conventional biochemical reactions into a few square centimeters of chips. The microfluidic chip based on the micro-droplets wraps single cells and reactants in the micro-droplets with picoliter capacity through a micron-sized channel to obtain a plurality of micro-reaction systems which are not interfered with each other, and analysis of the single cells is completed. As microfluidic prepared micro-droplets require few reagents to reach ultra-high throughput, they are of increasing interest in cell research.
After the microdroplet is generated, the microdroplet is further required to be captured and incubated, so that single cells wrapped in the microdroplet are fully combined with reactants. The micro droplet capturing and incubating devices used in the prior art are generally classified into an array type, a branched type, a height difference type, a serial slit type, and the like. The array device and the branch device are respectively designed with special upright posts or branch structures in the micro-channels, liquid drops are captured by utilizing the flow velocity and pressure changes in the micro-channels, and the liquid drops are enabled to be static in the device for incubation, however, the size of the upright posts in the array device needs to be matched with micro-liquid drops with corresponding sizes, universality is not achieved, and the micro-liquid drops captured by the branch device are limited by the device structures. The level difference device requires a complicated level change to be designed in the device, and slows down the flow rate of the micro-droplets to achieve the incubation purpose, however, this results in a complicated structure of the device itself and cannot be applied to micro-droplets with smaller size. The serial slit device captures the liquid drop through the channel matched with the shape of the liquid drop, so as to achieve the aim of incubation observation, however, the design of the channel needs to match the liquid drop with the corresponding size, and the device has no universality.
Disclosure of Invention
In order to solve at least one of the above shortcomings in the prior art, it is necessary to provide a microfluidic chip.
The application provides a microfluidic chip, which comprises a liquid drop inlet, a liquid drop outlet, a liquid drop incubation channel, a continuous phase channel and a confluence channel. The droplet incubation channel is connected to the droplet inlet for flowing the continuous phase and a plurality of droplets carried by the continuous phase into the droplet incubation channel. The continuous communication channel is positioned on at least one side of the droplet incubation channel and is communicated with the droplet incubation channel through an opening, so that the continuous phase enters the continuous phase channel through the opening after being separated from the droplet, and the droplet incubates in the droplet incubation channel. The continuous phase channel and the liquid drop incubation channel are intersected in a confluence channel, so that the continuous phase and the liquid drops after incubation are converged again at the confluence channel. The liquid drop outlet is connected with the confluence channel.
In some possible implementations, the number of continuous phase channels is two, with two continuous phase channels on opposite sides of the droplet incubation channel.
In some possible implementations, a direction along the droplet inlet to the droplet outlet is defined as a first direction, a direction from one continuous phase channel to another continuous phase channel is defined as a second direction, the continuous phase channels are divided into continuous phase bulk channels and continuous phase converging channels along the first direction, and a width of the continuous phase converging channels is smaller than a width of the continuous phase bulk channels along the second direction. The droplet incubation channel is divided into a droplet main body channel and a droplet converging channel along the first direction, and the width of the droplet converging channel is smaller than that of the droplet main body channel along the second direction. The continuous phase converging channel and the liquid drop converging channel meet at the converging channel.
In some possible implementations, the width of the droplet-converging channel decreases gradually along the first direction along the second direction.
In some possible implementations, an end of each continuous phase channel adjacent to the drop outlet is provided with a stopper for blocking a portion of the end of the continuous phase channel, and another portion of the end of the continuous phase channel that is not blocked forms the continuous phase converging channel.
In some possible implementations, the width of the end of the droplet body channel distal from the droplet converging channel increases gradually along the first direction.
In some possible implementations, a plurality of first posts are disposed between the continuous phase channel and the droplet incubation channel, and the opening is disposed between two adjacent first posts.
In some possible implementations, the first pillars are arranged in a plurality of rows, and the first pillars of two adjacent rows are staggered, so that the openings between the first pillars of two adjacent rows are staggered.
In some possible implementations, the microfluidic chip further includes a plurality of second pillars disposed in the continuous phase body channel.
In some possible implementations, the microfluidic chip further includes a mixing channel disposed on a side of the droplet inlet remote from the droplet incubation channel, the mixing channel including a plurality of curved portions and a plurality of straight portions, at least one end of each of the straight portions being connected to one of the curved portions.
According to the application, the continuous phase in the liquid drop incubation channel is separated from the liquid drop, so that the flow velocity of the liquid drop in the liquid drop incubation channel is reduced, the purposes of liquid drop incubation and observation are realized, the incubation time is accurate and controllable, and the problems of overlarge flow velocity and insufficient incubation time of the liquid drop in the liquid drop incubation channel caused by overlarge continuous phase flow are avoided. The application does not need to design complex height change in the device to slow down the flow velocity of the liquid drops, and also does not need to increase the transverse length of the liquid drop incubation channel to prolong the incubation time, thereby being beneficial to simplifying the chip structure. In addition, the application does not need to design a channel matched with the shape of the liquid drop to capture the liquid drop so as to achieve the aim of incubation observation, and the incubation process has no strict requirement on the size of the liquid drop and has universality.
Drawings
Fig. 1 is a schematic diagram of the overall structure of a microfluidic chip according to an embodiment of the present application.
Fig. 2 is a schematic diagram illustrating connection of a droplet generation module, a droplet incubation module, and a droplet sorting module in the microfluidic chip shown in fig. 1.
Fig. 3 is a schematic diagram of the droplet incubation module shown in fig. 2 in an embodiment.
Fig. 4 is a schematic cross-sectional view of the droplet incubation module shown in fig. 3 along A-A.
Fig. 5 is a schematic cross-sectional view of the droplet incubation module shown in fig. 3 along B-B.
Fig. 6 is a schematic cross-sectional view of the droplet incubation module shown in fig. 3 along C-C.
Fig. 7 is a schematic diagram of a first mixing channel of the droplet incubation module shown in fig. 2.
Fig. 8 is a schematic diagram of a droplet incubation module according to another embodiment of the application.
Fig. 9 is a partial schematic view of the droplet incubation module of fig. 8 at the first post and the opening.
Description of the main reference signs
Chip body 1
A substrate 1a
Cover plate 1b
Droplet generation module 10
A first continuous phase inlet 11
First dispersed phase inlet 12
A second dispersed phase inlet 13
First confluence passage 14
First droplet outlet 15
First electrode irrigation port 16
Second electrode casting opening 17
Droplet incubation module 20
First droplet inlet 21
Second droplet outlet 22
Droplet incubation channel 23
Second continuous phase channel 24
First upright 25
Second confluence passage 26
Second column 27
First mixing channel 28
Droplet sorting module 30
Second droplet inlet 31
A second continuous phase inlet 32
Third confluence passage 33
Drop collection outlet 34
Waste liquid drop outlet 35
Second mixing channel 36
Third electrode casting port 37
Fourth electrode irrigation port 38
Fifth electrode casting port 39
Sixth electrode casting port 39'
Third mixing channel 40
Microfluidic chip 100
First continuous phase channel 110
First dispersed phase channel 120
Second dispersed phase channel 130
First electrode runner 160
Second electrode runner 170
Droplet body channel 231
Droplet converging channel 232
Continuous phase body channel 241
Continuous phase convergence channel 242
Stop 243
Opening 250
Bending portion 281
Straight portion 282
Drop channel 310
Third continuous phase channel 320
Third electrode runner 370
Fourth electrode runner 380
First segment 2311
Second section 2312
Third section 2313
Fourth segment 2314
Fifth segment 2321
Sixth paragraph 2322
Included angle theta 1 、θ 2 、θ 3 、θ 4
Angle omega
Length L, L 1 、L 1a 、L 1b 、L 1c 、L 2 、L 3 、L 4 、L 5 、L 6 、L 7
Width W 1 、W 2 、W 3 、W 4 、W 5 、W 6 、W 7 、W 8 、W 9
Distance D 1 、D 2 、D 3 、D 4
Radius of radius R
The application will be further described in the following detailed description in conjunction with the above-described figures.
Detailed Description
In order to further describe the technical means and effects adopted by the present application to achieve the preset application, the following detailed description is made on the specific embodiments, structures, features and effects of the present application by referring to the accompanying drawings and the preferred embodiments.
Referring to fig. 1 and 2, an embodiment of the present application provides a microfluidic chip 100, which includes a chip body 1. The chip body 1 includes a base 1a and a cover plate 1b provided on the base 1 a. The substrate 1a is provided with a droplet generation module 10, a droplet incubation module 20 and a droplet sorting module 30. The droplet incubation module 20 is connected between the droplet generation module 10 and the droplet sorting module 30. In one embodiment, the droplet generation module 10, the droplet incubation module 20, and the droplet sorting module 30 are arranged in the same direction.
As shown in fig. 2, the droplet generation module 10 is used to generate droplets. In the present embodiment, the droplet generation module 10 includes a first continuous phase inlet 11, a first continuous phase channel 110, a first dispersed phase inlet 12, a first dispersed phase channel 120, a second dispersed phase inlet 13, a second dispersed phase channel 130, a first confluence channel 14, and a first droplet outlet 15. Wherein the first continuous phase inlet 11 is connected to the first continuous phase channel 110, the first dispersed phase inlet 12 is connected to the first dispersed phase channel 120, and the second dispersed phase inlet 13 is connected to the second dispersed phase channel 130. The first continuous phase channel 110, the first dispersed phase channel 120 and the second dispersed phase channel 130 converge in the first converging channel 14, and the first converging channel 14 is connected to the first droplet outlet 15.
The first continuous phase inlet 11 serves as an inlet for a continuous phase (e.g., oil phase) to enter the droplet-generating module 10, and the continuous phase enters the first continuous phase channel 110 through the first continuous phase inlet 11. The first dispersed phase inlet 12 serves as an inlet for a first dispersed phase (e.g., a reaction liquid, an aqueous phase) to enter the droplet generation module 10, and the first dispersed phase enters the first dispersed phase channel 120 through the first dispersed phase inlet 12. The second dispersed phase inlet 13 serves as an inlet for a second dispersed phase (e.g., biological sample, aqueous phase) to the droplet generation module 10, and the biological second dispersed phase enters the second dispersed phase channel 130 via the second dispersed phase inlet 13. The first dispersed phase and the biological second dispersed phase form a laminar flow at the junction, and then merge with the first continuous phase channel 110 into the first merging channel 14. In this case, the laminar flow is divided into a plurality of droplets by the shearing force of the continuous phase and is covered with the continuous phase by the interfacial tension between the two phases, for example, when the continuous phase is an oil phase and the dispersed phase is an aqueous phase, a plurality of water-in-oil microdroplets are formed. The first droplet outlet 15 is the outlet from the droplet generation module 10 as a droplet.
The biological sample is a biological fluid sample, and may be, for example, blood, plasma, serum, interstitial fluid, lymph fluid, urine, or the like of a human or other animal, or a culture fluid for culturing microorganisms or cells. The biological sample contains a target analyte (inclusion). The target analyte may be, but is not limited to, a protein, nucleic acid, liposome, peptide fragment, nucleotide, amino acid, virus, bacteria, parasite, cell, and some other single molecule or complex, etc.
Lysis is a critical step in obtaining target analytes within biological samples. The microfluidic chip 100 of the present application is subjected to cleavage using an electro-cleavage method. As shown in fig. 2, in one embodiment, the droplet-generating module 10 further comprises two first electrode irrigation ports 16 and two second electrode irrigation ports 17. Each first electrode molding opening 16 corresponds to one of the second electrode molding openings 17. The first electrode casting opening 16 and the second electrode casting opening 17 are respectively connected with the first electrode runner 160 and the second electrode runner 170, and the first electrode runner 160 and the second electrode runner 170 are respectively positioned at two opposite sides of the second dispersion channel 130.
One of the first electrode molding openings 16 is for inserting a first electrode metal strip, which may be a low melting point metal, which flows into the first electrode runner 160 after heating, thereby forming a first electrode (not shown) in the first electrode runner 160. The corresponding second electrode molding openings 17 are for inserting a second electrode metal strip, which may be a low melting point metal, which flows into the second electrode runner 170 after heating, thereby forming a second electrode (not shown) in the second electrode runner 170. The other first electrode casting opening 16 and the corresponding second electrode casting opening 17 serve as atmosphere communication openings for communicating with the atmosphere, thereby ensuring that the first electrode and the second electrode can be molded smoothly. One of the first electrode and the second electrode is a positive electrode, and the other is a negative electrode. The first electrode and the second electrode are used to generate an electric field for lysing the target analyte as the biological sample flows through the second dispersed phase channel 130. Thus, the subsequent reaction liquid can react with the target object to be detected in the biological sample to form a detection product carrying the detectable marker. For example, the reaction solution may comprise a lysis solution (e.g., lysozyme) which facilitates cell lysis, resulting in release of cell contents (e.g., nucleic acids), and a reaction base solution which may subsequently react with the lysed cell contents.
In one embodiment, the continuous phase has a flow rate of 0.5 to 20. Mu.L/min, the first dispersed phase has a flow rate of 0.05 to 1. Mu.L/min, and the second dispersed phase has a flow rate of 0.05 to 1. Mu.L/min. The particle size of the liquid drops is 15-100 mu m.
Referring to fig. 3, the droplet incubation module 20 includes a first droplet inlet 21, a second droplet outlet 22, a droplet incubation channel 23, two second continuous phase channels 24, and a second confluence channel 26. A droplet incubation channel 23 is located between the first droplet inlet 21 and the second droplet outlet 22. Two second continuous phase channels 24 are located on either side of the droplet incubation channel 23. The droplet incubation channels 23 are in communication with the two second continuous phase channels 24 through openings 250, respectively. In an embodiment, a plurality of first pillars 25 may be disposed between each of the second continuous phase channels 24 and the droplet incubation channels 23, and the openings 250 may be disposed between two adjacent first pillars 25.
Wherein the first droplet inlet 21 is connected to the first droplet outlet 15. Droplets exiting through the first droplet outlet 15 flow in from the first droplet inlet 21 and through the droplet incubation channel 23. Under the action of flow resistance, the disperse phase and the continuous phase are separated, specifically: the faster continuous phase will flow out of the droplet incubation channel 23 through opening 250 and into the second continuous phase channels 24 on both sides; at the same time, the slower flow rate of the dispersed phase (i.e., droplets including the first dispersed phase and the second dispersed phase) fails to pass through opening 250 to remain within droplet incubation channel 23 and flows along droplet incubation channel 23 under the remaining fluid pressure. Thus, the droplet flow rate within the droplet incubation channel 23 is reduced, facilitating droplet incubation and observation. Wherein, the incubation means that the target detection object in the liquid drop and the reaction base solution are uniformly mixed and connected in a certain incubation time. In one embodiment, the incubation time is 1 to 60 seconds.
Wherein, the droplet incubation module 20 is predefined to have a first direction, a second direction and a third direction perpendicular to each other. The first direction is a direction along the first droplet inlet 21 to the second droplet outlet 22, i.e., a length direction of the chip body 1. The second direction is a direction along one second continuous phase channel 24 to another second continuous phase channel 24, i.e., a width direction of the chip body 1. The third direction is the thickness direction of the chip body 1, i.e., the direction perpendicular to the paper surface, and is not labeled in fig. 2. It will be understood that the "length" of each section referred to below is the length in the first direction, the "width" of each section is the width in the second direction, and the "height" of each section is the height in the third direction.
In one embodiment, the width of the first droplet inlet 21 increases gradually along the first direction, and the angle θ between each side of the first droplet inlet 21 and the first direction 1 Is 5-30 degrees. Length L of each opening 250 2 Both (i.e. the distance between two adjacent first uprights 25) and the widthIs 5-50 μm so that the openings 250 can prevent the passage of droplets while allowing the continuous phase to pass. Length L of the plurality of first pillars 25 1 All the same, 20-100 μm.
In one embodiment, each second continuous phase channel 24 is divided into a continuous phase body channel 241 and a continuous phase converging channel 242 along the first direction. The cross section of the continuous phase body channel 241 defined along the first direction and the second direction may be set to be rectangular, and the width of the continuous phase converging channel 242 is smaller than the width of the continuous phase body channel 241. The droplet incubation channel 23 is divided into a droplet main body channel 231 and a droplet converging channel 232 in the first direction. The cross section of the droplet main body passage 231 defined in the first direction and the second direction may be set to be rectangular, and the width of the droplet converging passage 232 is smaller than the width of the droplet main body passage 231. The continuous phase converging channel 242 and the droplet converging channel 232 meet at a second converging channel 26, the second converging channel 26 connecting the second droplet outlet 22. In one embodiment, each second continuous phase channel 24 is provided with a stop 243 adjacent an end of the second droplet outlet 22. Stop 243 is used to block a portion of the end of the second continuous phase channel 24, and another portion of the end of the second continuous phase channel 24 that is not blocked forms continuous phase converging channel 242.
In one embodiment, the width of the droplet converging channel 232 gradually decreases along the first direction, and the angle ω of the droplet converging channel 232 itself is 15 to 60 degrees. The angle θ between adjacent sides of the continuous phase converging channel 242 and the droplet converging channel 232 2 (i.e., the angle of the stop 243 itself) is 35 to 80 degrees. The junction between the outer side of the second confluence channel 26 and the outer side of each second continuous phase channel 24 is smoothly transited, and the radius R of the junction is 50-200 μm.
Referring to fig. 4 to 6, in one embodiment, the width W of the first droplet inlet 21 1 Width W of continuous phase body channel 241 is 100-200 μm 2 400-1200 μm width W of continuous phase converging channel 242 3 50-200 μm, width W of first pillar 25 4 5-50 μm. Width W of droplet body channel 231 5 100 to 400 μm, the length L (see FIG. 3) of the droplet main body channel 231 is6000 to 10000 μm, and the height of the droplet main body channel 231 is 5 to 50 μm. Spacing D between continuous phase converging channel 242 and droplet converging channel 232 1 200-400 mu m. Width W of droplet converging channel 232 6 100-200 mu m.
In this way, the droplets in the droplet main body passage 231 can be arranged in a certain order (single row arrangement) without binding up when flowing to the droplet converging passage 232. The continuous phase and the incubated droplets re-converge at the junction. The second droplet outlet 22 serves as an outlet for droplets exiting the droplet incubation module 20.
Since the cross section of the droplet main body passage 231 defined in the first direction and the second direction is rectangular, the width of the droplet main body passage 231 is substantially the same in the first direction. In another embodiment, the cross-sections of the continuous phase body channel 241 and the droplet body channel 231 are not limited to rectangular, and may be changed according to actual needs. As shown in fig. 8 and 9, the width of the end of the droplet main body channel 231 distant from the droplet converging channel 232 gradually increases in the first direction, specifically, the droplet main body channel 231 is defined to be divided into a first segment 2311, a second segment 2312, a third segment 2313, and a fourth segment 2314 in this order in the first direction, and the width of the first segment 2311 gradually increases in the first direction. Thus, when a droplet flows from the first droplet inlet 21 into the droplet body channel 231, the continuous phase can separate from the droplet more quickly and enter the second continuous phase channels 24 on both sides via the openings 250, avoiding droplet packing. In the present embodiment, the angle θ between the first segment 2311 and the first direction 3 45 degrees or less. Further, the width of the first droplet inlet 21 may also be gradually reduced along the first direction, in which case the angle θ between each side of the first droplet inlet 21 and the first direction 1 0-20 degrees, thereby avoiding accumulation of droplets at dead corner positions on both sides with the flow rate of droplets just entering the droplet body channel 231 kept almost unchanged.
Further, in order to reduce the flow rate of the continuous phase in the opening 250, and avoid the situation that a part of droplets are carried through the opening 250 and blocked when the flow rate of the continuous phase is too high, the first pillars 25 are further configured in a multi-row structure, so as to increase the number of channels for the continuous phase to flow between the droplet main channels 231 and the continuous phase main channels 241, i.e. the flow rate of the dispersed continuous phase. That is, the plurality of first pillars 25 of each row are aligned in the first direction, and the plurality of rows of first pillars 25 are stacked in the second direction. The first posts 25 of the first, second and third segments 2311, 2312, 2313 are illustrated in fig. 8 and 9 as having three rows and the first post 25 of the fourth segment 2314 is illustrated as having two rows, however, the number of rows of the first posts 25 is not limited by the present application. Wherein, the first pillars 25 of two adjacent rows are staggered, so that the openings 250 of two adjacent rows are also staggered. In this way, the continuous phase in the continuous phase main body channel 241 is blocked by the second row of the first pillars 25 when flowing out through the openings 250 of the first row of the first pillars 25 adjacent to the droplet main body channel 231, and flows out to the openings 250 on both sides of the first pillars 25, and the continuous phase flows out through the openings 250 of the second row of the first pillars 25, similarly, and thus the continuous phase flow rate is dispersed.
In the present embodiment, for any one of the first, second, third and fourth segments 2311, 2312, 2313 and 2314, the distance D of the first pillars 25 of the adjacent two rows in the second direction 3 Distance D between two adjacent first uprights 25 of each row 4 The same applies to the range of 5 to 50. Mu.m. Length L of first plurality of posts 25 in first section 2311 1 The same, 20-100 μm, length L of first segment 2311 3 200-1000 μm. Length L of first plurality of posts 25 in second section 2312 1 The same, 20-100 μm, length L of second segment 2312 4 800-1600 mu m. Length L of each first post 25 in the first row in third section 2313 1a 40-200 μm, greater than the length L of the first upright posts 25 of other rows 1 Length L of third segment 2313 5 600-2000 μm. Length L of first column 25 of the first row of fourth segment 2314 1b 200-1000 μm, greater than the length L of the second row of first pillars 25 1c And L 1 The length of the fourth segment 2314 is the total length L of the continuous phase body channel 241 and the length L of the first segment 2311 3 Length L of second segment 2312 4 And length L of third segment 2313 5 And (3) a difference. Wherein the length L 1b And length L 1c Is L as the difference of 1 . In the second row of fourth segment 2314,length L 1c And a length of L 1 The first columns 25 of (1) are alternately arranged, L 1c >L 1 。
That is, the length of first row of first posts 25 of first section 2311 and second section 2312 is minimal. Thus, when a droplet flows from the first droplet inlet 21 into the droplet main body passage 231 of the preceding stage, the continuous phase is separated from the droplet more quickly and enters the second continuous phase passages 24 on both sides through the openings 250. As the droplets flow in the droplet-body channel 231, the continuous-phase content of the droplet-body channel 231 becomes smaller, and thus the lengths of the first-row first pillars 25 at which the third and fourth segments 2313 and 2314 may be disposed relatively increase.
In addition, in order to disperse the incubated droplets, the configuration of the droplet collecting channel 232 may be changed so that the distance between adjacent droplets is controlled within a range that enables the subsequent droplet sorting module 30 to sort independently. As shown in fig. 9, the width of the droplet-converging channel 232 is gradually reduced in the first direction and then is uniform, i.e., the droplet-converging channel 232 is divided into a fifth segment 2321 and a sixth segment 2322 in the first direction. The width of the fifth segment 2321 decreases gradually in the first direction, the length L of the fifth segment 2321 6 100-200 mu m. The width of sixth segment 2322 remains constant along the first direction, length L of sixth segment 2322 7 Width W of sixth segment 2322 is 100-200 μm 8 Matching the size of the liquid drop after incubation, the liquid drop is 20-60 μm. Each side of fifth segment 2321 is angled at θ from each side of sixth segment 2322 4 The angle θ between each side of the end of the continuous phase converging channel 242 and the first direction is also 4 ，θ 4 110-150 degrees. Width W of continuous phase converging channel 242 9 100-200 mu m. According to the droplet incubation module 20 disclosed by the application, the continuous phase in the droplet incubation channel 23 is separated from the droplet, so that the flow rate of the droplet in the droplet incubation channel 23 is reduced, and the purposes of droplet incubation and observation are realized, wherein the length of the droplet incubation channel 23 is fixed, the flow rate of the droplet is reduced, the incubation time is accurate and controllable, and the problems of overlarge flow rate of the droplet in the droplet incubation channel 23 and insufficient incubation time caused by overlarge flow rate of the continuous phase are avoided. The application does not require complex height variations in the device design to slow down the dropletNor does it need to increase the lateral length of the droplet incubation channel 23 to extend the incubation time, which is advantageous for simplifying the chip structure. In one embodiment, the height of the chip body 1 is 10-50 μm. In addition, the application does not need to design a channel matched with the shape of the liquid drop to capture the liquid drop so as to achieve the aim of incubation observation, and the incubation process has no strict requirement on the size of the liquid drop and has universality. In addition, after incubation, the separated continuous phase can be converged with the incubated liquid drops again, so that the waste of materials is avoided, and the preparation cost is saved.
Referring again to fig. 1 and 3, in one embodiment, the droplet incubation module 20 further includes a plurality of second posts 27 disposed in the continuous phase body channel 241. The second column 27 is fixed between the bottom of the continuous phase body channel 241 and the lower surface of the cover plate 1b for supporting in the continuous phase body channel 241. Further, the plurality of second pillars 27 in each continuous phase body channel 241 may be arranged at intervals along the first direction, and the distance D between two adjacent second pillars 27 2 200-400 mu m. The cross-sectional shape of each second pillar 27 along the extension plane of the chip body 1 may be provided in a regular polygon or a circle. As shown in fig. 5, the width W of each second pillar 27 7 50-200 mu m.
Referring to fig. 2 and fig. 7 together, in an embodiment, the droplet incubation module 20 further includes a first mixing channel 28 disposed between the first droplet outlet 15 and the first droplet inlet 21. The first mixing channel 28 includes a plurality of curved portions 281 and a plurality of straight portions 282, with at least one end of each straight portion 282 having one curved portion 281 attached thereto. For example, the first blending channel 28 may be a serpentine curve. In this way, the first mixing channel 28 can continuously change the flow direction of the droplets flowing out through the first droplet outlet 15, so that the droplets flow in a single row under the action of inertia to reduce aggregation into a cluster. Specifically, the droplets flow from the left side into the first mixing channel 28, against the side of the largest radius when encountering the first bend 281, along the side of the smallest radius when encountering the second bend 281, along the side of the largest radius when arriving at the third bend 281, and so on. The straight portion 282 is used for pulling the distance between the front and rear drops, so that the effect of orderly single-row flowing of the drops is enhanced.
Referring again to fig. 2, the droplet sorting module 30 includes a second droplet inlet 31, a droplet channel 310, a second continuous phase inlet 32, a third continuous phase channel 320, a third converging channel 33, a droplet collection outlet 34, and a waste droplet outlet 35. Wherein the second droplet inlet 31 is connected to the droplet channel 310 and the second continuous phase inlet 32 is connected to the third continuous phase channel 320. The droplet passage 310 and the third continuous phase passage 320 converge on the third converging passage 33. The droplet collection outlet 34 and the discard droplet outlet 35 are connected to the third confluence passage 33.
Wherein the second droplet inlet 31 is connected to the second droplet outlet 22. The liquid droplets flowing out through the second liquid droplet outlet 22 flow in from the second liquid droplet inlet 31. The second continuous phase inlet 32 is used as an inlet for a continuous phase (such as an oil phase) to enter the droplet sorting module 30, and the continuous phase enters the third continuous phase channel 320 through the second continuous phase inlet 32, and after being intersected with the droplets, the droplets are driven to be transported in the third confluence channel 33. In one embodiment, the second continuous phase inlet 32 communicates with a power generation device (not shown). The power generation device is used for generating power, so that the continuous phase is led into the third continuous phase channel 320 under the action of the power and drives the liquid drops to be transported. The power generating device may be a negative pressure generator or a positive pressure generator.
In one embodiment, the drop sorting module 30 further includes a second mixing channel 36 disposed between the second drop inlet 31 and the second drop outlet 22. The second mixing channel 36 may have a similar structure to the first mixing channel 28 for a neat single row flow of droplets exiting through the second droplet outlet 22 to reduce agglomeration.
Wherein, the separation is a key step after obtaining the liquid drops, and is used for extracting the liquid drops (hereinafter referred to as liquid drops to be detected) with the target detection objects with the expected properties from all the liquid drops. And droplets containing undesired products or lacking the target detection object (hereinafter referred to as waste droplets) need to be removed by sorting. In one embodiment, the drop sorting module 30 further includes a third electrode molding port 37 and a fourth electrode molding port 38. The third electrode molding port 37 and the fourth electrode molding port 38 are connected to the third electrode runner 370 and the fourth electrode runner 380, respectively, and the third electrode runner 370 and the fourth electrode runner 380 are located at opposite sides of the third confluence channel 33, respectively.
The third electrode molding port 37 is used for inserting a third electrode metal strip, which flows into the third electrode runner 370 after being heated, thereby forming a third electrode (not shown) in the third electrode runner 370. The fourth electrode molding opening 38 is used to insert a fourth electrode metal strip, which flows into the fourth electrode runner 380 after being heated, thereby forming a fourth electrode (not shown) in the fourth electrode runner 380. One of the third electrode and the fourth electrode is a positive electrode, and the other is a negative electrode. The third electrode and the fourth electrode are used for generating an electric field, when the liquid drops flow through the third confluence channel 33, the electric field is used for deflecting the micro liquid drops, the liquid drops to be detected in the liquid drops enter the liquid drop collecting outlet 34 after deflection, and the waste liquid drops in the liquid drops enter the waste liquid drop outlet 35 after deflection, so that the sorting purpose is realized.
Specifically, a droplet signal in the droplet may be collected by a droplet signal collecting device (such as a high-speed camera, a fluorescence detector, and a raman spectrum detector, not shown), and the droplet may be determined to be a droplet to be detected or a waste micro droplet according to the collected droplet signal. When it is determined that a droplet to be measured, the electric field between the third electrode and the fourth electrode is used to deflect the droplet to the droplet collection outlet 34; when it is determined that a droplet is discarded, the electric field between the third electrode and the fourth electrode is used to deflect the droplet to the discarded droplet outlet 35. In one embodiment, the flow rate of the second continuous phase is 1 to 20. Mu.L/min, such that when the second continuous phase drives the transport of droplets, the distance between two adjacent droplets is greater than the minimum distance required for sorting by 200. Mu.m.
In one embodiment, the drop sorting module 30 further includes a fifth electrode molding port 39 and a sixth electrode molding port 39'. The fifth electrode molding port 39 and the sixth electrode molding port 39' are connected to a fifth electrode runner (not shown) and a sixth electrode runner (not shown), respectively, which are located at opposite sides of the third confluence channel 33, respectively. The fifth and sixth electrode runners may be located before or after the third and fourth electrode runners 370 and 380.
The fifth electrode pouring opening is used for inserting a fifth electrode metal strip, and the fifth electrode metal strip flows into the fifth electrode runner after being heated, so that a fifth electrode is formed in the fifth electrode runner. The sixth electrode pouring opening is used for inserting a sixth electrode metal strip, and the sixth electrode metal strip flows into the sixth electrode runner after being heated, so that a sixth electrode is formed in the sixth electrode runner. One of the fifth electrode and the sixth electrode is a positive electrode, and the other is a negative electrode. The fifth electrode and the sixth electrode serve as shielding electrodes for avoiding droplet accumulation at the front end under the influence of the deflecting electric field.
As shown in fig. 2, in one embodiment, the drop sorting module 30 further includes a third mixing channel 40 disposed before the drop collection outlet 34 and the discard drop outlet 35. The third mixing channel 40 may have a similar structure to the first mixing channel 28 for a neat single row flow of deflected droplets to reduce agglomeration into a cluster.
The various inlets and outlets in the present application are provided on the surface of the chip body 1. The various channels are located inside the chip body 1. In an embodiment, the various inlets and outlets may be located on the same surface of the chip body 1, i.e. on the surface of the substrate 1a or the surface of the cover plate 1b.
The application integrates the droplet generation module 10, the droplet incubation module 20 and the droplet sorting module 30 on the same chip to form an integrated chip, so that the cross contamination problem between biological samples and the instability problem of fluid, which may be caused by the integration of a plurality of chips, are avoided.
Although the present application has been described with reference to the preferred embodiments, it should be understood that the present application is not limited to the specific embodiments, and that various changes and modifications can be made by one skilled in the art without departing from the scope of the application.
Claims (10)
1. The microfluidic chip is characterized by comprising a liquid drop inlet, a liquid drop outlet, a liquid drop incubation channel, a continuous phase channel and a confluence channel;
the liquid drop incubation channel is connected with the liquid drop inlet and is used for allowing a continuous phase and a plurality of liquid drops carried by the continuous phase to flow into the liquid drop incubation channel;
the continuous communication channel is positioned on at least one side of the droplet incubation channel and is communicated with the droplet incubation channel through an opening, so that the continuous phase enters the continuous phase channel through the opening after the continuous phase is separated from the droplet, and the droplet is incubated in the droplet incubation channel;
the continuous phase channel and the liquid drop incubation channel are intersected in a confluence channel, so that the continuous phase and the liquid drops after incubation are converged again at the confluence channel;
the liquid drop outlet is connected with the confluence channel.
2. The microfluidic chip of claim 1, wherein the number of continuous phase channels is two, two of the continuous phase channels being located on opposite sides of the droplet incubation channel.
3. The microfluidic chip according to claim 2, wherein a direction from said droplet inlet to said droplet outlet is defined as a first direction, a direction from one of said continuous phase channels to the other of said continuous phase channels is defined as a second direction, said continuous phase channels are divided into a continuous phase bulk channel and a continuous phase converging channel along said first direction, and a width of said continuous phase converging channel is smaller than a width of said continuous phase bulk channel along said second direction;
the liquid drop incubation channel is divided into a liquid drop main body channel and a liquid drop converging channel along the first direction, and the width of the liquid drop converging channel is smaller than that of the liquid drop main body channel along the second direction;
the continuous phase converging channel and the liquid drop converging channel meet at the converging channel.
4. The microfluidic chip of claim 3, wherein the width of said droplet-converging channel decreases gradually along said first direction.
5. The microfluidic chip according to claim 3, wherein an end portion of each of said continuous phase channels adjacent to said droplet outlet is provided with a stopper for blocking a part of said end portion of said continuous phase channel, and another part of said end portion of said continuous phase channel which is not blocked forms said continuous phase converging channel.
6. The microfluidic chip of claim 3, wherein the width of the end of the droplet body channel distal from the droplet converging channel increases gradually in the first direction.
7. The microfluidic chip according to claim 1, wherein a plurality of first posts are disposed between the continuous phase channel and the droplet incubation channel, and the openings are disposed between two adjacent first posts.
8. The microfluidic chip according to claim 7, wherein said first pillars are arranged in a plurality of rows, and said first pillars of adjacent rows are staggered such that said openings between said first pillars of adjacent rows are staggered.
9. The microfluidic chip of claim 1, further comprising a plurality of second pillars disposed in the continuous phase body channel.
10. The microfluidic chip of claim 1, further comprising a mixing channel disposed on a side of said droplet inlet remote from said droplet incubation channel, said mixing channel comprising a plurality of curved portions and a plurality of straight portions, at least one end of each of said straight portions being connected to one of said curved portions.
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