CN218321413U - Digital micro-fluidic chip for nucleic acid detection - Google Patents
Digital micro-fluidic chip for nucleic acid detection Download PDFInfo
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- CN218321413U CN218321413U CN202222650811.9U CN202222650811U CN218321413U CN 218321413 U CN218321413 U CN 218321413U CN 202222650811 U CN202222650811 U CN 202222650811U CN 218321413 U CN218321413 U CN 218321413U
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Abstract
The utility model discloses a digital micro-fluidic chip for nucleic acid detection, which comprises an upper polar plate and a lower polar plate; the lower pole plate is fixed below the upper pole plate, and a first cavity is formed between the lower pole plate and the upper pole plate; an electrode array and a fluorescence detection hole are arranged in the first cavity, the electrode array is arranged on the upper surface of the lower polar plate, and the fluorescence detection hole is arranged in the electrode array; the electrode array comprises a plurality of driving electrodes and a liquid storage electrode, and the liquid storage electrode is connected to the fluorescence detection hole through the driving electrodes; the sample to be detected required by the detection sample liquid storage tank required by the microfluidic chip is extremely trace, so that the cost of nucleic acid detection is greatly reduced; the micro-fluidic chip is small in size, a detection sample is convenient to add, the liquid movement is controlled by the electrodes completely, the mixing process does not need manual operation, and the operation difficulty is greatly reduced.
Description
Technical Field
The utility model relates to a liquid measurement field especially relates to a digital micro-fluidic chip for nucleic acid detects.
Background
Nucleic acid detection is one of the most important detection means in clinical detection at present. With the development of scientific technology and the improvement of highly automated detection technology, the development of automated, high-throughput and rapid nucleic acid detection technology is greatly promoted, and the method has great significance for preventing the spread of infectious diseases, early screening of diseases and accelerating the laboratory experiment progress.
The existing nucleic acid detection method still stays in a real-time fluorescence quantitative PCR method, and has the defects of complex operation, long consumed time, slow reaction, large reagent and sample consumption, high detection cost and the like. In addition, the real-time fluorescence quantitative PCR method requires precise temperature setting, so that the dependence on expensive instruments limits the application range, and the problems of long time consumption, complex steps, heavy equipment, high cost and the like in nucleic acid detection can be caused.
SUMMERY OF THE UTILITY MODEL
To the above problem, the utility model provides a digital micro-fluidic chip and detection method for nucleic acid detects mainly solves the problem among the background art.
The utility model provides a digital micro-fluidic chip for nucleic acid detection, which comprises an upper polar plate and a lower polar plate; the lower polar plate is fixed below the upper polar plate, and a first cavity is formed between the lower polar plate and the upper polar plate; an electrode array and a fluorescence detection hole are arranged in the first cavity, the electrode array is arranged on the upper surface of the lower polar plate, and the fluorescence detection hole is arranged in the electrode array; the electrode array comprises a plurality of driving electrodes and liquid storage electrodes, and the liquid storage electrodes are connected to the fluorescence detection holes through the driving electrodes.
The further improvement is that the lower polar plate comprises a substrate layer, a hydrophobic layer and a dielectric layer, the electrode array is paved on the surface of the substrate layer at intervals, the dielectric layer covers the electrode array, the hydrophobic layer is arranged above the dielectric layer, and the dielectric layer is made of photoresist.
The further improvement is that the substrate layer is a glass substrate layer covered with a chromium coating having a pattern of electrode arrays formed using positive photoresist, the electrode arrays being formed by etching chromium nitrate from the chromium coating on the glass.
The further improvement is that the fluorescence detection hole is a pure transparent round hole which is not covered with metal chromium in the middle.
The further improvement is that the hydrophobic layer of the lower polar plate is a Teflon hydrophobic layer.
The further improvement is that the upper polar plate is made of glass materials, and a layer of indium tin oxide film and a hydrophobic layer formed by Teflon are covered on one surface of the upper polar plate.
The further improvement is that the upper polar plate and the lower polar plate are connected by three layers of double-sided adhesive tapes to form a first cavity, and the gap is filled with silicon oil.
The further improvement lies in that a plurality of liquid storage tanks are further arranged in the first cavity, liquid storage tanks are provided with liquid storage ports, the liquid storage ports are arranged in gaps at the edge of the lower polar plate, and the liquid storage tanks are arranged above the liquid storage electrodes.
The liquid storage tanks comprise a constant-temperature amplification mixed liquid storage tank, a gene editing nucleic acid detection liquid storage tank, a detection sample storage tank and a waste liquid tank, and the constant-temperature amplification mixed liquid storage tank, the gene editing nucleic acid detection liquid storage tank, the detection sample storage tank and the waste liquid tank are communicated by a driving electrode.
The further improvement is that the driving electrode and the liquid storage electrode are connected to the upper polar plate through copper wires, and the copper wires penetrate through the upper surface of the upper polar plate to form a contact array.
Compared with the prior art, the utility model discloses a digital micro-fluidic chip for nucleic acid testing has following advantage at least:
1) The amount of the sample detection liquid storage tank required by the microfluidic chip is extremely trace, so that the cost of nucleic acid detection is greatly reduced;
2) The micro-fluidic chip has small volume, is convenient to add detection liquid, does not need to be operated by multiple persons, controls the liquid to move by electrodes, does not need manual operation in the mixing process, and greatly reduces the operation difficulty;
3) The dielectric layer uses photoresist as material, in order to produce the liquid drop drive phenomenon and separate the electrode and liquid contact;
4) The gap between the upper and lower plates is filled with silicone oil, which surrounds the droplets to facilitate droplet operation and prevent evaporation;
5) The contact array is convenient for the digital microfluidic chip to be connected into the socket of the equipment, and the driving electrode and the liquid storage electrode are started to carry out liquid operation so as to move liquid.
Drawings
The drawings are for illustrative purposes only and are not to be construed as limiting the patent; for the purpose of better illustrating the embodiments, certain features of the drawings may be omitted, enlarged or reduced, and do not represent the size of an actual product; it will be understood by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted.
Fig. 1 is a schematic view of the overall structure of an embodiment of the present invention;
fig. 2 is a schematic view of an electrode structure according to an embodiment of the present invention;
fig. 3 is a schematic diagram of a layered structure of a microfluidic chip according to an embodiment of the present invention.
Reference numerals
1. An upper polar plate; 2. a lower polar plate; 3. a constant temperature amplification mixed liquid storage tank; 4. a gene editing nucleic acid detection liquid storage tank; 5. detecting a sample liquid storage tank; 6. a waste liquid tank; 7. a fluorescence detection well; 8. a contact array; 9. a drive electrode; 10. a reservoir electrode; 11. a base layer; 12. a dielectric layer; 13. a hydrophobic layer; 14. an upper electrode glass plate; 15. an indium tin oxide film; 16. a silicone oil.
Detailed Description
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted" and "connected" are to be interpreted broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, so to speak, as communicating between the two elements. The specific meaning of the above terms in the present invention can be understood as specific cases to those skilled in the art. The technical solution of the present invention will be further explained with reference to the accompanying drawings and examples.
Referring to fig. 1, a digital microfluidic chip for nucleic acid detection comprises an upper plate 1 and a lower plate 2, wherein the lower plate 2 is fixed below the upper plate 1, and a first cavity is formed between the lower plate 2 and the upper plate 1; an electrode array and a fluorescence detection hole 7 are arranged in the first cavity, the electrode array is arranged on the upper surface of the lower polar plate 2, and the fluorescence detection hole 7 is arranged in the electrode array; the electrode array comprises a plurality of driving electrodes 9 and liquid storage electrodes 10, and the liquid storage electrodes 10 are connected to the fluorescence detection holes 7 through the driving electrodes 9; the sample to be detected of the sample liquid storage tank 5 required by the micro-fluidic chip is extremely micro, so that the cost of nucleic acid detection is greatly reduced; the micro-fluidic chip is small in size, a detection sample is convenient to add, the micro-fluidic chip does not need to be operated by multiple persons, the liquid is controlled to move by the electrodes completely, manual operation is not needed in the mixing process, and the operation difficulty is greatly reduced.
Specifically, the liquid storage electrode 10 is arranged close to the edge of the lower polar plate 2, so that liquid can be added conveniently, and the operation difficulty is reduced.
Specifically, the driving electrode 9 and the liquid storage electrode 10 are connected to the upper polar plate 1 through copper wires, the copper wires penetrate through the upper surface of the upper polar plate 1 to form a contact array 8, the contact array 8 is convenient for the digital microfluidic chip to be connected into an equipment socket, and the driving electrode 9 and the liquid storage electrode 10 are started to carry out liquid operation so that liquid moves.
Specifically, the area of the liquid storage electrode 10 is larger than that of the driving electrode 9, so that after various liquids are loaded into the digital microfluidic chip, the volume of the stored liquid is far larger than that of the driving electrode 9 taken out once, the liquids can be taken out from the liquid storage electrode 10 for multiple times, and the adding times of the liquids of the liquid storage electrode 10 are reduced.
Specifically, the reservoir electrode 10 is 5.4 × 3.6mm in size and can absorb about 7ul of fluid when energized, and the driver electrode 9 is 2.2 × 2.2mm in size and can absorb about 2ul of fluid when energized.
As a preferred embodiment of the present invention, the bottom plate 2 includes a substrate layer 11, a hydrophobic layer 13 and a dielectric layer 12, the electrode array is spread on the surface of the substrate layer 11 at intervals, the electrode array is covered with the dielectric layer 12, the hydrophobic layer 13 is arranged above the dielectric layer 12, and the material of the dielectric layer 12 is photoresist.
As a preferred embodiment of the present invention, the substrate layer 11 is a glass substrate layer, and the glass substrate layer is covered with a chromium coating layer, and the chromium coating layer is formed by etching a chromium coating layer on glass with a pattern electrode array formed using a positive photoresist.
As a preferred embodiment of the present invention, the fluorescence detection hole 7 is a pure transparent circular hole without covering chromium metal in the middle.
Specifically, the fluorescence detection hole 7 is a pure transparent circular hole which is not covered with metal chromium in the middle and is used for detecting a fluorescence signal, so that the result of the reaction is judged to be negative or positive.
As a preferred embodiment of the utility model, the hydrophobic layer 13 of bottom plate 2 is teflon hydrophobic layer 13, and upper plate 1 is the glass material, and the single face covers the hydrophobic layer 13 that one deck indium tin oxide membrane 15 and teflon formed, and spray teflon on upper plate 1, bottom plate 2 can reduce the adhesion of detection sample on the electrode, reduces the resistance of droplet motion, is favorable to reducing the possibility of cross contamination between the sample simultaneously.
As a preferred embodiment of the utility model, connect by three-layer double-sided tape between upper plate 1 and the lower plate 2, form first cavity, and fill the gap with silicon oil 16, three-layer double-sided tape provides the space and prevents that upper and lower plate from sliding for the liquid drop flows, silicon oil 16 surrounds the liquid drop, in order to promote the liquid drop operation and prevent to evaporate, the clearance of first cavity is little, the required sample volume of awaiting measuring of the required detection sample liquid storage pond of micro-fluidic chip is extremely micro, the cost of nucleic acid detection has greatly been reduced.
Specifically, the distance between the upper plate 1 and the lower plate 2 is preferably 360um.
As a preferred embodiment of the utility model, still be equipped with a plurality of liquid storage ponds in the first cavity, the liquid storage pond is equipped with the stock solution mouth, and the stock solution mouth sets up in the gap at 2 edges of polar plate down, and the liquid storage pond position sets up in stock solution electrode 10 tops.
Specifically, the liquid storage tanks comprise a constant-temperature amplification mixed liquid storage tank 3, a gene editing nucleic acid detection liquid storage tank 4, a detection sample storage tank 5 and a waste liquid tank 6, and liquid storage electrodes 10 and driving electrodes 9 communicated with the liquid storage tanks are connected below the liquid storage tanks; the liquid storage electrode 10 and the driving electrode 9 are responsible for accumulation of electric charges and electric field gradients, and when a voltage is applied, the liquid storage electrode 10 and the driving electrode 9 are activated, and the liquid drop is manipulated by changing the electric charges and the electric field gradients along the electrode wires, so that the movement of the liquid drop is controlled to complete the step of detecting the nucleic acid.
As the utility model discloses a preferred embodiment, for filling in the first cavity has protective gas, avoids receiving external environmental pollution.
The operation process of nucleic acid detection in the embodiment of the present invention is;
the method comprises the following steps: the isothermal amplification mixture was withdrawn by syringe and injected into the isothermal amplification mixture reservoir 3, which in this example was 5.4 x 3.6mm in size, and was capable of adsorbing about 7 μ L of fluid when energized.
Step two: the gene-editing nucleic acid detecting solution was extracted by a syringe and injected into the gene-editing nucleic acid detecting solution reservoir 4, and in this example, about 7. Mu.L of the solution was adsorbed after energization, and the size was 5.4X 3.6mm.
Step three: the sample to be tested is drawn out by means of a syringe and injected into the test sample reservoir 5, in this example, about 7 μ L of liquid can be adsorbed after power-on, with a size of 5.4 x 3.6mm.
Step four: after the power is on, when the electrodes are activated, 2 μ L of the sample to be detected and 2 μ L of the isothermal amplification mixed solution are respectively conveyed to the central position of the chip through the liquid drops formed by the driving electrodes 9 to be repeatedly mixed, so that the isothermal amplification is fully reacted.
Step five: after the reaction is finished, the liquid drop is controlled to be divided into two parts, one part is controlled to be conveyed to the waste liquid pool 6 through the driving electrode 9, and the other part is continuously remained at the central position of the original chip.
Step six: next, 2. Mu.L of the gene-editing nucleic acid detection solution was controlled to be fed to the center of the chip, and mixed with the above solution to carry out a detection reaction.
Step seven: after the reaction is finished, the liquid drop is moved to the fluorescence detection hole 7 for fluorescence signal detection.
It will be appreciated that the syringe is injected through the gap between the upper and lower plates 2 through the silicone oil 16 to ensure that the liquid does not come into contact with the outside air, reducing the error of the test.
It should be understood that the principle of controlling the droplet to have one second is that the droplet is 4 μ L, which is larger than the volume of one grid of the driving electrodes 9, so that the droplet occupies the area of the grid part of the other driving electrodes 9 at the periphery, and when the driving electrodes 9 on both sides of the droplet are simultaneously activated, the suction force at both ends is simultaneously generated to divide the droplet into two.
It should be understood that the central position refers to the area of fig. 1 where the four electrodes surround the fluorescence detection hole 7, and the repeated mixing refers to the movement of the liquid droplet in the circumferential direction around the fluorescence detection hole 7, and during the movement, various components in the liquid droplet are continuously mixed due to external force, and finally, a uniform liquid droplet is formed.
Specifically, after the liquid in the waste liquid tank 6 is fully stored, the waste liquid in the waste liquid tank 6 is extracted through the needle cylinder to be drained.
In the drawings, the positional relationship is described for illustrative purposes only and is not to be construed as limiting the present patent; it is obvious that the above embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. This need not be, nor should it be exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (10)
1. A digital microfluidic chip for nucleic acid detection, comprising:
an upper polar plate; and
the lower pole plate is fixed below the upper pole plate, and a first cavity is formed between the lower pole plate and the upper pole plate;
an electrode array and a fluorescence detection hole are arranged in the first cavity, the electrode array is arranged on the upper surface of the lower polar plate, and the fluorescence detection hole is arranged in the electrode array;
the electrode array comprises a plurality of driving electrodes and a liquid storage electrode, and the liquid storage electrode is connected to the fluorescence detection hole through the driving electrodes.
2. The digital microfluidic chip for nucleic acid detection according to claim 1, wherein the bottom plate comprises a substrate layer, a hydrophobic layer and a dielectric layer, the electrode array is laid on the surface of the substrate layer at intervals, the electrode array is covered with the dielectric layer, the hydrophobic layer is arranged above the dielectric layer, and the dielectric layer is made of photoresist.
3. The digital microfluidic chip for nucleic acid detection according to claim 2, wherein the substrate layer is a glass substrate layer covered with a chromium coating having a pattern of electrode arrays formed using positive photoresist, the electrode arrays formed by etching chromium coatings on glass with chromium nitrate.
4. The digital microfluidic chip for nucleic acid detection according to claim 3, wherein the fluorescence detection hole is a pure transparent circular hole without metal chromium covering the middle.
5. The digital microfluidic chip for nucleic acid detection according to claim 2, wherein the hydrophobic layer of the lower plate is a teflon hydrophobic layer.
6. The digital microfluidic chip for nucleic acid detection according to claim 1, wherein the upper plate is made of glass, and a single surface of the upper plate is covered with a layer of indium tin oxide film and a hydrophobic layer formed by teflon.
7. The digital microfluidic chip for nucleic acid detection according to claim 1, wherein the upper plate and the lower plate are connected by a three-layer double-sided tape to form a first cavity, and the gap is filled with silicone oil.
8. The digital microfluidic chip for nucleic acid detection according to claim 7, wherein a plurality of liquid reservoirs are further disposed in the first cavity, the liquid reservoirs are provided with liquid storage ports, the liquid storage ports are disposed in the gaps at the edge of the lower plate, and the liquid reservoirs are disposed above the liquid storage electrodes.
9. The digital microfluidic chip for nucleic acid detection according to claim 8, wherein the reservoirs comprise a constant temperature amplification mixed solution reservoir, a gene editing nucleic acid detection solution reservoir, a detection sample reservoir and a waste solution reservoir, and the constant temperature amplification mixed solution reservoir, the gene editing nucleic acid detection solution reservoir, the detection sample reservoir and the waste solution reservoir are connected by a driving electrode.
10. The digital microfluidic chip for nucleic acid detection according to claim 1, wherein the driving electrode and the reservoir electrode are connected to the upper plate through copper wires, and the copper wires penetrate through the upper surface of the upper plate to form a contact array.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202222650811.9U CN218321413U (en) | 2022-09-30 | 2022-09-30 | Digital micro-fluidic chip for nucleic acid detection |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202222650811.9U CN218321413U (en) | 2022-09-30 | 2022-09-30 | Digital micro-fluidic chip for nucleic acid detection |
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| Publication Number | Publication Date |
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| CN218321413U true CN218321413U (en) | 2023-01-17 |
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| CN202222650811.9U Active CN218321413U (en) | 2022-09-30 | 2022-09-30 | Digital micro-fluidic chip for nucleic acid detection |
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