CN112588332B - Micro-droplet generation method and generation system - Google Patents
Micro-droplet generation method and generation system Download PDFInfo
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Abstract
The application provides two micro-droplet generation systems and a micro-droplet generation method adopting the two micro-droplet generation systems. The micro-droplet generation system comprises a micro-fluidic chip and an electrode driving unit; the micro-fluidic chip comprises a top cover and an electrode layer, wherein the top cover comprises an upper cover, a conductive layer and a first hydrophobic layer which are sequentially arranged, the electrode layer comprises a second hydrophobic layer, a dielectric layer and an electrode array layer which are sequentially arranged, the first hydrophobic layer and the second hydrophobic layer are oppositely arranged, and a liquid drop channel layer is formed between the first hydrophobic layer and the second hydrophobic layer. The large droplets are controlled to pass through the electrode array layer, and the electrodes are operated to enable the large droplets to leave small droplets on the path through which the large droplets pass. Or array hydrophilic modification is carried out on the upper cover, and when a large liquid drop passes through the hydrophilic point, a small liquid drop is left at the hydrophilic point due to the hydrophilic action of the hydrophilic point. Compared with the traditional method capable of generating small liquid drops through digital microfluidics, the two micro-liquid drop generation systems and the method thereof can greatly shorten the liquid drop generation time.
Description
Technical Field
The invention relates to the technical field of droplet control, in particular to a method and a system for generating micro droplets.
Background
How to uniformly decompose a certain volume of liquid into a large number of droplets with uniform volumes is one of the key problems to be solved in the microfluidic technology, and is a key link in application fields including digital polymerase chain reaction (ddPCR), digital loop-mediated isothermal amplification (dLAMP), digital enzyme-linked immunosorbent assay (dELISA), unicellular omics and the like. The current technical means for generating nanoliter liquid drops in high flux mainly comprise a micro-droplet micro-fluidic technology and a micro-well micro-fluidic technology. Representative microfluidic droplet flow technologies include Bio-Rad and 10 XMenomics, and the technology is characterized in that a high-precision micropump is used for controlling oil, and a cross-shaped structure is used for continuously extruding sample liquid so as to generate a large number of small droplets from picoliters to nanoliters. The method for generating nano-liter droplets at high flux based on the micro-droplet micro-fluidic technology relies on the accurate control of the pressure of a high-precision micro pump and the high-precision chip processing technology based on MEMS, the generated droplets are still stored in the same container, each droplet needs to be detected one by one through a micro channel during detection, the equipment cost is high, and the system is complex. Microwell microfluidics is typified by Thermo Fisher, which is a technique characterized by mechanically spreading a sample solution over an array of microwells such that the sample is evenly distributed into each microwell, forming small droplets on the order of picoliters. The technology based on micro-well micro-fluidic usually needs to uniformly coat the reagent on the surface of the micro-well array by means of mechanical force, and then fill the upper surface and the lower surface of the micro-well with inert medium liquid.
Digital microfluidics, due to its ability to manipulate each droplet independently, is another technology for high throughput droplet generation, and WO 2016/170109 Al and US20200061620A1 both describe a method for generating a large number of droplets based on a digital microfluidic platform. However, the method for generating nanoliter droplets at high throughput based on digital microfluidic technology described in the above patent mainly uses digital microfluidic technology to manipulate a large droplet to generate a small droplet, and then transport the small droplet to a corresponding position. The main disadvantage of this method is the slow droplet generation and the long sample preparation time.
Disclosure of Invention
In view of the above, it is desirable to provide a method and a system for generating micro-droplets with a fast droplet generating speed and a stable and controllable droplet generating speed.
A micro-droplet generation system comprises a micro-fluidic chip and an electrode driving unit;
the microfluidic chip comprises a top cover and an electrode layer, wherein the top cover comprises an upper cover, a conducting layer and a first hydrophobic layer which are sequentially arranged, the electrode layer comprises a second hydrophobic layer, a dielectric layer and an electrode array layer which are sequentially arranged, the first hydrophobic layer and the second hydrophobic layer are oppositely arranged, and a liquid drop channel layer is formed between the first hydrophobic layer and the second hydrophobic layer;
the electrode driving unit is connected with the electrode array layer, and the electrode driving unit controls the opening or closing of the electrodes of the electrode array layer, so that large droplets added to the droplet channel layer are controlled to flow in the droplet channel layer, and micro droplets are formed at a plurality of preset positions of the droplet channel layer.
In one embodiment, the volume of the formed microdroplets is determined by the number of electrodes that are open at a preset position.
A micro-droplet generation system comprises a micro-fluidic chip and an electrode driving unit;
the microfluidic chip comprises a top cover and an electrode layer, wherein the top cover comprises an upper cover, a conducting layer and a first hydrophobic layer which are sequentially arranged, the electrode layer comprises a second hydrophobic layer, a dielectric layer and an electrode array layer which are sequentially arranged, the first hydrophobic layer and the second hydrophobic layer are oppositely arranged, and a liquid drop channel layer is formed between the first hydrophobic layer and the second hydrophobic layer;
through hydrophilic modification, a hydrophilic dot array is formed on one side, away from the conducting layer, of the first hydrophobic layer, and at least one electrode is arranged between every two adjacent hydrophilic dots at intervals;
the electrode driving unit is connected with the electrode array layer, the electrode driving unit is used for driving large liquid drops to flow in the liquid drop flow channel layer, and the large liquid drops form micro liquid drops at the hydrophilic points.
In one embodiment, the microdroplet-forming volume is determined by the area of the hydrophilic dot.
In one embodiment, the electrodes of the electrode array layer are hexagonal in shape.
A method for generating micro-droplets by using the micro-droplet generating system is characterized by comprising the following steps:
and controlling the opening or closing of the electrodes of the electrode array layer, so that micro droplets are formed at a plurality of preset positions of the electrode array layer when the large droplets flow through the electrode array layer.
In one embodiment, the plurality of preset positions are the same or different in size so as to simultaneously generate microdroplets of the same or different volumes.
In one embodiment, the operation of controlling the opening or closing of the electrodes of the electrode array layer to make the large droplets flow through the electrode array layer to form micro droplets at a plurality of preset positions of the electrode array layer is as follows:
s110, opening the electrodes from the first row to the P-th row, and adding the solution to the electrodes from the first row to the P-th row of the electrode array layer to form large liquid drops, wherein P is a positive integer;
s120, keeping the electrodes at the first preset position of the first row open, closing other electrodes of the first row, simultaneously opening the electrodes of the P +1 th row, driving the large liquid drops to move forward one row on the electrode array layer, forming micro liquid drops at the first preset position of the first row, and at least 1 electrode is arranged between the adjacent first preset positions;
s130, keeping the electrodes at the second preset position of the second row open, closing other electrodes of the second row, simultaneously, opening the electrodes of the P +2 th row, driving the large liquid drops to move forward one row on the electrode array layer, forming micro liquid drops at the second preset position of the second row, at least 1 electrode is arranged between every two adjacent second preset positions at intervals, and the first preset position and the second preset position are in different rows;
s140, keeping the electrode at the nth preset position of the nth row open, closing other electrodes of the nth row, simultaneously opening the electrode of the P + nth row, driving the large liquid drop to move forward one row on the electrode array layer, forming a micro-liquid drop at the nth preset position of the nth row, at least spacing 1 electrode between the adjacent nth preset positions, and enabling the nth-1 preset position and the nth preset position to be in different columns, wherein n is a positive integer greater than 3;
and S150, repeatedly executing S140, and forming a plurality of micro droplets on the electrode array layer until the large droplets are exhausted.
In one embodiment, the volume of the microdroplets is controlled by adjusting the distance between the first hydrophobic layer and the second hydrophobic layer and the size of the single electrode.
A method for producing a droplet of the above-described droplet production system, comprising the steps of:
and controlling the opening or closing of the electrodes of the electrode array layer, so that when the large liquid drop flows through the electrode array layer, a micro liquid drop is formed at the hydrophilic point array of the electrode array layer.
In one embodiment, the operation of controlling the opening or closing of the electrodes of the electrode array layer to form micro droplets at the hydrophilic dot array of the electrode array layer when the large droplets flow through the electrode array layer is as follows:
s210, opening the electrodes from the first row to the P-th row, and adding the solution to the electrodes from the first row to the P-th row of the electrode array layer to form large liquid drops, wherein P is a positive integer;
s220, closing the electrodes in the first row, simultaneously opening the electrodes in the P +1 th row, driving the large liquid drops to move forward one row in the electrode array layer, and forming micro liquid drops at the hydrophilic point position of the first row;
s230, closing the electrodes in the second row, simultaneously opening the electrodes in the P +2 th row, driving the large liquid drops to move forward one row on the electrode array layer, and forming micro liquid drops at the hydrophilic point position of the second row;
s240, closing the electrode of the nth row, meanwhile, opening the electrode of the P + nth row, driving the large liquid drop to move forward one row on the electrode array layer, and forming a micro liquid drop at the hydrophilic point of the nth row, wherein n is a positive integer larger than 3;
and S250, repeatedly executing S240, and forming a plurality of micro droplets on the electrode array layer until the large droplets are exhausted.
In one embodiment, the volume of the microdroplets is controlled by controlling the size of the hydrophilic dots.
The two micro-droplet generation systems and the corresponding micro-droplet generation methods are different from the traditional method for generating small droplets by digital microfluidics, and the traditional digital microfluidics generates a small droplet by controlling a large droplet and then conveys the small droplet to a corresponding position. In the two micro-droplet generation methods, the large droplet is controlled to pass through the electrode array layer, and the small droplet is left on the path of the large droplet by operating the electrodes. Or array hydrophilic modification is carried out on the upper cover, and when a large liquid drop passes through the hydrophilic point, a small liquid drop is left at the hydrophilic point due to the hydrophilic action of the hydrophilic point. Compared with the traditional method for generating small liquid drops by digital microfluidics, the two micro-drop generation methods can greatly shorten the time for generating the liquid drops.
Drawings
Fig. 1 is a schematic cross-sectional view of a microfluidic chip according to an embodiment;
FIG. 2 is a schematic structural diagram of a microfluidic chip according to another embodiment;
FIG. 3 is a flow chart of a method for generating micro-droplets using the microfluidic chip shown in FIG. 1;
FIG. 4 is a schematic diagram of a process for forming a micro-droplet by moving a large droplet;
FIG. 5 is a schematic flow chart of the movement of a large droplet to form a plurality of micro droplets;
FIG. 6 is a schematic flow chart of an embodiment of a large droplet moving on an electrode array layer to form a plurality of micro droplets;
FIG. 7 is a schematic diagram of another embodiment of a large droplet moving on an electrode array layer to form a plurality of micro droplets;
fig. 8 is a flowchart of a method of generating micro-droplets using the microfluidic chip shown in fig. 2.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In the description of the present invention, it should be understood that the relation indicating the orientation or position such as "above" is based on the orientation or position relation shown in the drawings, or the orientation or position relation which the product of the present invention is usually put into use, or the orientation or position relation which is usually understood by those skilled in the art, and is only for convenience of describing the present invention and simplifying the description, but does not indicate or imply that the device or element to be referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, cannot be construed as limiting the present invention.
Furthermore, the terms "first," "second," and the like are used merely to distinguish one description from another, and are not to be construed as indicating or implying relative importance.
The following detailed description of embodiments of the invention refers to the accompanying drawings.
One embodiment of a micro-droplet generation system includes a microfluidic chip and an electrode driving unit (not shown) as shown in fig. 1.
The micro-fluidic chip comprises a top cover and an electrode layer, wherein the top cover comprises an upper cover 5, a conducting layer 6 and a first hydrophobic layer 7 which are sequentially arranged. The electrode layer comprises a second hydrophobic layer 9, a dielectric layer 10 and an electrode array layer 11 which are arranged in sequence. The first hydrophobic layer 7 and the second hydrophobic layer 9 are arranged opposite to each other, and a droplet channel layer is formed between the first hydrophobic layer 7 and the second hydrophobic layer 9.
The electrode driving unit is connected with the electrode array layer 11, and the electrode driving unit controls the large droplets added to the droplet flow channel layer to flow in the droplet flow channel layer by controlling the opening or closing of the electrodes of the electrode array layer 11, so that micro droplets are formed at the preset position of the droplet flow channel layer. It will be appreciated that the plurality of predetermined locations may be the same or different sizes to simultaneously generate the same or different volumes of microdroplets.
In this embodiment, the shape of the electrodes is hexagonal. When the electrode shape is hexagonal, the contact surface becomes large, and the utilization rate of the electrode plate is higher. It will be appreciated that the shape of the electrodes may also be any other shape or combination of shapes.
According to the micro-droplet generation system, the large droplets are added into the droplet flow channel layer 8, the electrodes of the electrode array layer 11 are controlled to be opened or closed through the electrode driving unit, so that the large droplets added into the droplet flow channel layer 8 are controlled to flow on the surface of the electrode array layer 11 in a manner similar to coating, micro-droplets are formed at a plurality of preset positions of the droplet flow channel layer 8, the droplet generation time can be greatly shortened, the droplet generation stability is improved, the size of generated droplets can be dynamically adjusted according to requirements, and the operation process is simple and convenient. And equipment such as a high-precision micro pump and the like is not needed, so that the system cost is reduced. And the expansion capability is strong, and more small droplets or a plurality of groups of samples can be separated by expanding the microfluidic size.
The present application provides another embodiment of a micro-droplet generation system, comprising a microfluidic chip and an electrode driving unit (not shown) as shown in fig. 2.
The microfluidic chip comprises a top cover 13 and an electrode layer 14, wherein the top cover 13 comprises an upper cover, a conductive layer and a first hydrophobic layer which are sequentially arranged, the electrode layer comprises a second hydrophobic layer, a dielectric layer and an electrode array layer which are sequentially arranged, the first hydrophobic layer and the second hydrophobic layer are oppositely arranged, and a droplet channel layer is formed between the first hydrophobic layer and the second hydrophobic layer (the layer structure diagram 2 of the microfluidic chip is not shown, and reference can be made to fig. 1).
Referring to fig. 2, by hydrophilic modification, a hydrophilic dot array is formed on a side of the first hydrophobic layer away from the conductive layer, and at least one electrode is spaced between adjacent hydrophilic dots 12. The electrode driving unit is connected with the electrode array layer, the electrode driving unit is used for driving the large liquid drops to flow in the liquid drop flow channel layer, and the large liquid drops form micro liquid drops at the hydrophilic points. It will be appreciated that the above-described microdroplet generation system, forming microdroplet volumes, is determined by the hydrophilic dot area.
According to the micro-droplet generation system, the large droplets are added into the droplet flow channel layer, the electrode driving unit is used for driving the large droplets to flow in the droplet flow channel layer, and when the large droplets pass through the hydrophilic points 12, the small droplets are left at the hydrophilic points due to the hydrophilic action of the hydrophilic points, so that the droplet generation time can be greatly shortened. In addition, the micro-droplet generation system does not need to separate small droplets through a control electrode, so that the operation is simpler and more convenient. And equipment such as a high-precision micro pump and the like is not needed, so that the system cost is reduced. And the expansion capability is strong, and more small droplets or a plurality of groups of samples can be separated by expanding the microfluidic size.
The present application also provides an embodiment of a method for generating micro-droplets of the micro-droplet generating system shown in fig. 1, including the following steps:
and controlling the opening or closing of the electrodes of the electrode array layer, so that when the large liquid drops flow through the electrode array layer, micro liquid drops are formed at a plurality of preset positions of the electrode array layer respectively.
According to the micro-droplet generation method, the opening or closing of the electrodes of the electrode array layer is controlled, so that when large droplets flow through the electrode array layer, micro-droplets are formed at a plurality of preset positions of the electrode array layer respectively. Can greatly shorten the generation time of the liquid drops and has simple and convenient operation flow.
It will be appreciated that the plurality of predetermined locations may be the same size or different sizes to simultaneously generate different volumes of microdroplets.
Furthermore, at least one electrode is arranged between the preset positions at intervals. The plurality of preset positions are spaced apart from each other by at least one electrode to avoid droplet coalescence. Preferably, the plurality of predetermined positions are spaced apart from each other by 2 electrodes.
Specifically, referring to fig. 3, the operation of controlling the opening or closing of the electrodes of the electrode array layer to form micro droplets at a plurality of preset positions of the electrode array layer when the large droplets flow through the electrode array layer is as follows:
s110, opening the electrodes from the first row to the P-th row, and adding the solution to the electrodes from the first row to the P-th row of the electrode array layer to form large liquid drops, wherein P is a positive integer.
S120, keeping the electrodes at the first preset position of the first row open, closing other electrodes of the first row, simultaneously, opening the electrodes of the P +1 th row, driving the large liquid drops to move forward one row on the electrode array layer, forming micro liquid drops at the first preset position of the first row, and at least spacing 1 electrode between the adjacent first preset positions.
S130, keeping the electrodes at the second preset positions of the second row open, closing other electrodes of the second row, simultaneously, opening the electrodes of the P +2 th row, driving the large liquid drops to move one row forward on the electrode array layer, forming micro liquid drops at the second preset positions of the second row, at least 1 electrode is arranged between every two adjacent second preset positions at intervals, and the first preset positions and the second preset positions are in different rows.
S140, keeping the electrode at the nth preset position of the nth row open, closing other electrodes of the nth row, simultaneously opening the electrodes of the P + nth row, driving the large liquid drop to move forward one row on the electrode array layer, forming a micro liquid drop at the nth preset position of the nth row, at least spacing 1 electrode between adjacent nth preset positions, and enabling the nth-1 preset position and the nth preset position to be in different columns, wherein n is a positive integer greater than 3.
And S150, repeatedly executing S40, and forming a plurality of micro droplets on the electrode array layer until the large droplets are exhausted.
It is understood that the specific operations of S140 repeatedly executed in S150 are: n is 3, execute S140 once; n is 4, execute S140 once; n is 5, and S140, \ 8230is performed once until the large droplet is exhausted. That is, the large droplets move in the first row to the nth row in sequence, and a plurality of micro droplets are formed in each of the first row to the nth row.
It will be appreciated that the "rows" in the above described method of microdroplet generation may be indicated by "columns". That is, the large droplets move in the first to nth rows in sequence, and a plurality of micro droplets are formed in each of the first to nth rows.
In one embodiment, the volume of the micro-droplets is controlled by adjusting the distance between the first hydrophobic layer 7 and the second hydrophobic layer 9 and the size of the individual electrodes. The droplet volume can be accurately controlled between picoliters and microliters by adjusting the distance between the first hydrophobic layer 7 and the second hydrophobic layer 9 and the size of the individual electrodes.
Specifically, referring to fig. 4, an electrode array composed of electrodes 1 controls the large droplet 2 to move along the electrode array in the direction of the arrow on the way. The separation of a droplet 3 from a bulk liquid 2 is achieved by controlling the electrode array. The large drop 2 continues to move in the direction of the arrow while the small drop 3 remains in place. As further shown in FIG. 5, the large droplet 2 is made to leave a plurality of small droplets 3 on its moving path by repeating the operation shown in FIG. 4, the small droplets 3 are separated by a plurality of electrodes to prevent the small droplets from combining, the electrodes below the small droplets 3 are opened to fix the small droplets 3 in place, and the separation step is stopped or repeated until the large droplets are completely consumed after the target number of small droplets can be separated. As further shown in fig. 6, the large droplets 4 are manipulated according to steps S1 to S6 to leave a plurality of small droplets 3 on the path, the small droplets 3 are separated by a plurality of electrodes to avoid the small droplets from combining, the electrodes under the small droplets 3 are opened to fix the small droplets 3 in place, and the separation step is stopped or repeated until the large droplets are completely consumed after the target number of small droplets can be separated. A cross-sectional view of a small droplet 3 is shown in fig. 1, the droplet being located between a first hydrophobic layer 77 and a second hydrophobic layer 99. The volume of the small droplets 3 can be accurately controlled between picoliters and microliters by adjusting the gap 8 and the size of the electrodes 11.
Referring to fig. 7, when the electrodes are different in size or one to several adjacent electrodes are turned on simultaneously, droplets of different sizes may be formed on the electrode array layer.
The present application also provides an embodiment of a method for generating microdroplets by using the microdroplet generation system shown in fig. 2, comprising the following steps:
and controlling the opening or closing of the electrodes of the electrode array layer, so that when the large liquid drop flows through the electrode array layer, a micro liquid drop is formed at the hydrophilic point array of the electrode array layer.
In one embodiment, the volume of the microdroplets is controlled by controlling the size of the hydrophilic dots.
According to the micro-droplet generation method, the large droplets are added into the droplet flow channel layer, the electrode driving unit is used for driving the large droplets to flow in the droplet flow channel layer, and when the large droplets pass through the hydrophilic points 12, the small droplets are left at the hydrophilic points 12 due to the hydrophilic action of the hydrophilic points 12, so that the droplet generation time can be greatly shortened. In addition, the micro-droplet generation system does not need to separate small droplets through a control electrode, so that the operation is simpler and more convenient.
Referring to fig. 8, when the electrodes of the electrode array layer are controlled to be turned on or off to allow a large droplet to flow through the electrode array layer, the formation of a micro-droplet at the hydrophilic dot array of the electrode array layer is performed as follows:
s210, opening the electrodes from the first row to the P-th row, and adding the solution to the electrodes from the first row to the P-th row of the electrode array layer to form large liquid drops, wherein P is a positive integer;
s220, closing the electrodes in the first row, simultaneously opening the electrodes in the P +1 th row, driving the large liquid drops to move forward one row in the electrode array layer, and forming micro liquid drops at the hydrophilic point positions of the first row;
s230, closing the electrodes in the second row, simultaneously opening the electrodes in the P +2 th row, driving the large liquid drops to move forward one row on the electrode array layer, and forming micro liquid drops at the hydrophilic point position of the second row;
s240, closing the electrode of the nth row, simultaneously opening the electrode of the P + nth row, driving the large liquid drop to move forward one row on the electrode array layer, and forming a micro liquid drop at the hydrophilic point of the nth row, wherein n is a positive integer larger than 3;
and S250, repeatedly executing S240, and forming a plurality of micro droplets on the electrode array layer until the large droplets are exhausted.
It can be understood that the specific operations of repeatedly performing S240 in S250 are: n is 3, execute S240 once; n is 4, execute S240 once; n is 5, and S240, \ 8230is performed once until the large droplet is exhausted. That is, the large droplets move in the first row to the nth row in sequence, and a plurality of micro droplets are formed in each of the first row to the nth row.
It will be appreciated that the "rows" in the above described method of microdroplet generation may be indicated by "columns". That is, the large droplets move in the first to nth rows in sequence, and a plurality of micro droplets are formed in each of the first to nth rows.
In the two methods for generating the micro-droplets, the separation steps are repeated to separate the droplets with target quantity.
The two methods for generating the micro-droplets are different from the traditional method for generating the small-droplets by digital microfluidics, and the traditional method for generating the small-droplets by controlling the large-droplet to transport the small-droplet to the corresponding position. In the two micro-droplet generation methods, the large droplet is controlled to pass through the electrode array layer, and the small droplet is left on the path of the large droplet by operating the electrodes. Or array hydrophilic modification is carried out on the upper cover, and when a large liquid drop passes through the hydrophilic point, a small liquid drop is left at the hydrophilic point due to the hydrophilic action of the hydrophilic point. Compared with the traditional method for generating small liquid drops by digital microfluidics, the two micro-drop generation methods can greatly shorten the time for generating the liquid drops.
According to the two micro-droplet generation methods, the control similar to coating is realized by driving the large droplets on the electrode array layer, and the generation of droplets with high flux and nanoliter level can be realized by controlling the electrodes or performing array hydrophilic modification on the upper cover. The volume of the liquid drop can be accurately adjusted by adjusting the size of the electrode, the gap distance of the electrode, the size of the hydrophilic modification point and the like. After the separation of the high-flux nano-liter liquid drops is finished, corresponding experiments and detection can be carried out on the digital microfluidic chip. The method can realize biochemical application functions such as ddPCR, dLAMP, dELISA single cell experiments and the like by matching with an optical detection module. Can be applied to other nucleic acid detection such as isothermal amplification. Meanwhile, any small liquid drop of the chip can be screened or independently tested, and more small liquid drops or a plurality of groups of samples can be separated by expanding the size of the chip.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (7)
1. A micro-droplet generation method adopting a micro-droplet generation system, wherein the micro-droplet generation system comprises a micro-fluidic chip and an electrode driving unit; the micro-fluidic chip includes top cap and electrode layer, the top cap is including upper cover, conducting layer and the first hydrophobic layer that sets gradually, the electrode layer is including the second hydrophobic layer, dielectric layer and the electrode array layer that set gradually, first hydrophobic layer with the second hydrophobic layer sets up relatively, first hydrophobic layer with form the liquid drop runner layer between the second hydrophobic layer, its characterized in that, the method includes:
the electrode driving unit is connected with the electrode array layer and controls the electrodes of the electrode array layer to be opened or closed, so that when large liquid drops flow through the electrode array layer, micro liquid drops are formed at a plurality of preset positions of the electrode array layer respectively; the method comprises the following steps:
s110, opening electrodes from the first row to the P-th row, and adding a solution to the electrodes from the first row to the P-th row of the electrode array layer to form the large liquid drops, wherein P is a positive integer;
s120, keeping the electrodes at the first preset position of the first row open, closing other electrodes of the first row, simultaneously opening the electrodes of the P +1 th row, driving the large liquid drops to move forward one row on the electrode array layer, forming micro liquid drops at the first preset position of the first row, and at least 1 electrode is arranged between the adjacent first preset positions;
s130, keeping the electrodes at the second preset position of the second row open, closing other electrodes of the second row, simultaneously, opening the electrodes of the P +2 th row, driving the large liquid drops to move forward one row on the electrode array layer, forming micro liquid drops at the second preset position of the second row, at least 1 electrode is arranged between every two adjacent second preset positions at intervals, and the first preset position and the second preset position are in different rows;
s140, keeping an electrode at an nth preset position of an nth row open, closing other electrodes of the nth row, simultaneously opening electrodes of a P + nth row, driving a large liquid drop to move forward one row on the electrode array layer, forming a micro liquid drop at the nth preset position of the nth row, at least 1 electrode is arranged between adjacent nth preset positions at intervals, and the nth-1 preset position and the nth preset position are in different rows, wherein n is a positive integer greater than 3;
and S150, repeatedly executing S140, and forming a plurality of micro droplets on the electrode array layer until the large droplets are exhausted.
2. A method of producing microdroplets as claimed in claim 1 wherein the volume of microdroplets formed is determined by the number of electrodes that are open at predetermined positions.
3. The method of claim 2, wherein the plurality of predetermined locations are the same or different in size to simultaneously generate the same or different volumes of microdroplets.
4. A method of generating microdroplets as claimed in claim 3 wherein the volume of the microdroplet is controlled by adjusting the distance between the first hydrophobic layer and the second hydrophobic layer and the size of the individual electrodes.
5. A micro-droplet generation method adopting a micro-droplet generation system, wherein the micro-droplet generation system comprises a micro-fluidic chip and an electrode driving unit; the micro-fluidic chip includes top cap and electrode layer, the top cap is including upper cover, conducting layer and the first hydrophobic layer that sets gradually, the electrode layer is including the second hydrophobic layer, dielectric layer and the electrode array layer that set gradually, first hydrophobic layer with the second hydrophobic layer sets up relatively, first hydrophobic layer with form the liquid drop runner layer between the second hydrophobic layer, its characterized in that, the method includes:
through hydrophilic modification, a hydrophilic dot array is formed on one side, away from the conducting layer, of the first hydrophobic layer, and at least one electrode is arranged between every two adjacent hydrophilic dots at intervals;
the electrode driving unit is connected with the electrode array layer and controls the opening or closing of the electrodes of the electrode array layer, so that when large liquid drops flow through the electrode array layer, micro liquid drops are formed at the hydrophilic point array of the electrode array layer; the method comprises the following steps:
s210, opening the electrodes from the first row to the P-th row, and adding the solution to the electrodes from the first row to the P-th row of the electrode array layer to form the large liquid drop, wherein P is a positive integer;
s220, closing the electrodes in the first row, simultaneously opening the electrodes in the P +1 th row, driving the large liquid drops to move forward one row in the electrode array layer, and forming micro liquid drops at the hydrophilic point position of the first row;
s230, closing the electrodes in the second row, simultaneously opening the electrodes in the P +2 th row, driving the large liquid drops to move forward one row on the electrode array layer, and forming micro liquid drops at the hydrophilic point position of the second row;
s240, closing the electrode of the nth row, meanwhile, opening the electrode of the P + nth row, driving the large liquid drop to move forward one row on the electrode array layer, and forming a micro liquid drop at the hydrophilic point of the nth row, wherein n is a positive integer larger than 3;
and S250, repeatedly executing S240, and forming a plurality of micro droplets on the electrode array layer until the large droplets are exhausted.
6. A method of microdroplet generation as claimed in claim 5 wherein the volume of the microdroplet is controlled by controlling the size of the hydrophilic dot.
7. The method of claim 6, wherein the electrodes of the electrode array layer are hexagonal in shape.
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