CN117987250A - Microfluidic chip - Google Patents
Microfluidic chip Download PDFInfo
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- CN117987250A CN117987250A CN202410113889.8A CN202410113889A CN117987250A CN 117987250 A CN117987250 A CN 117987250A CN 202410113889 A CN202410113889 A CN 202410113889A CN 117987250 A CN117987250 A CN 117987250A
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- base plate
- microfluidic chip
- cover plate
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- 238000001514 detection method Methods 0.000 claims abstract description 29
- 239000000758 substrate Substances 0.000 claims abstract description 26
- 238000006243 chemical reaction Methods 0.000 claims abstract description 23
- 239000007788 liquid Substances 0.000 claims abstract description 17
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 13
- 230000003321 amplification Effects 0.000 claims abstract description 12
- 238000002347 injection Methods 0.000 claims abstract description 9
- 239000007924 injection Substances 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims description 10
- 230000009471 action Effects 0.000 claims description 9
- 238000005452 bending Methods 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 239000002313 adhesive film Substances 0.000 claims description 3
- 210000001124 body fluid Anatomy 0.000 claims description 3
- 239000010839 body fluid Substances 0.000 claims description 3
- 238000007599 discharging Methods 0.000 claims description 3
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 3
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 3
- 239000002699 waste material Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 2
- 238000007789 sealing Methods 0.000 abstract description 3
- 238000012864 cross contamination Methods 0.000 abstract description 2
- 238000013461 design Methods 0.000 abstract description 2
- 238000005192 partition Methods 0.000 abstract description 2
- 238000009827 uniform distribution Methods 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract 1
- 238000007397 LAMP assay Methods 0.000 description 10
- 238000005516 engineering process Methods 0.000 description 8
- 230000006872 improvement Effects 0.000 description 8
- 238000011160 research Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000012123 point-of-care testing Methods 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000006820 DNA synthesis Effects 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
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- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 238000012544 monitoring process Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
The invention discloses a microfluidic chip, which comprises a substrate and a cover plate arranged at the upper end of the substrate in a sealing manner, wherein a micro-channel for flowing in and out of liquid from the upper end face of the cover plate from one side of the substrate is arranged between the substrate and the upper cover plate, and the micro-channel comprises a sample injection area, a reaction area, a temperature control area, a mixing area and a detection area. The invention has simple structure and reasonable design, can realize the uniform distribution of samples in one sample inlet to different reaction chambers, realize the simultaneous amplification of multiple target partitions of one sample inlet, can prevent the cross contamination between different reaction tanks, reduce the operation of repeated sample injection of operators, and realize the detection with high sensitivity, simplicity and stability.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a microfluidic chip.
Background
Loop-mediated isothermal amplification (LAMP) is a common nucleic acid amplification technology capable of rapidly and efficiently amplifying target DNA or RNA sequences at constant temperature. The basic principle of the LAMP technique is that a plurality of primers and a specific DNA polymerase are combined to generate a continuous DNA synthesis reaction on a target sequence, thereby forming a characteristic loop structure. These loop structures can be generated continuously during the reaction and visualized after the amplification reaction. Compared with the traditional Polymerase Chain Reaction (PCR), the method has the following characteristics of isothermal reaction, rapidness, high efficiency, high specificity, direct visualization and the like. In a word, the LAMP technology is used as a simple, rapid and efficient nucleic acid amplification method, is widely applied to biological research and practical application in various fields, and provides a powerful tool for rapid diagnosis, monitoring and research.
The microfluidic chip (Microfluidic Chip) is a micro laboratory that integrates micro fluidic processing elements such as micro channels, mixers, separators, control valves, and detectors. It achieves accurate control and operation of minute volumes of liquid on a small-sized chip. The design of microfluidic chips is based on microfluidic principles, using micro-scale channels and structures to manipulate and process micro-samples, typically operating in the nano-liter to microliter liquid volume range. Advantages of microfluidic chips include: small volume and quick reaction, precise control, multifunctional integration, low cost and high efficiency.
The combination of the microfluidic chip technology and the LAMP technology can meet the multiple detection requirements of the LAMP technology and simultaneously reduce the problem of LAMP reaction aerosol pollution; meanwhile, the combination of the two can greatly expand the detection application range of the LAMP technology and meet the demand of point-of-CARE TESTING (POCT) detection. In summary, the microfluidic chip technology and the LAMP detection method are combined, so that a specific, stable, sensitive, rapid and efficient instant detection system can be established, and the method has important application value in the aspects of pathogen detection in the fields of medicine, food, environment, pets and the like, and is expected to solve the defects of time consumption, labor consumption and high cost in detection.
Many researches at present combine the LAMP technology with a microfluidic chip to realize simple and rapid detection. However, the researches still need auxiliary equipment for providing constant temperature at different points such as multiple sample inlets, a water bath kettle and the like. Cannot be detected in a non-laboratory environment and requires the operation of a professional, and home-based detection cannot be achieved.
Disclosure of Invention
The present invention is directed to a microfluidic chip for solving the problems set forth in the background art.
In order to achieve the above purpose, the present invention provides the following technical solutions: the micro-fluidic chip comprises a substrate and a cover plate arranged at the upper end of the substrate in a sealing manner, wherein a micro-channel used for flowing in and out of one side of the substrate from the upper end face of the cover plate is arranged between the substrate and the upper cover plate, and the micro-channel comprises a sample injection area, a reaction area, a temperature control area, a mixing area and a detection area.
As a further improvement of the present invention, the sample injection area includes:
the first pool groove is arranged on the upper end face of the cover plate;
The first round groove is arranged on the lower end surface of the cover plate and is communicated with the first pool groove;
the second circular groove is arranged at the upper end of the substrate and is matched with the first circular groove, and the first circular groove and the second circular groove are respectively arranged in an array at intervals with the horizontal plane, are identical in size and are vertically aligned in sequence.
As a further improvement of the present invention, the reaction zone comprises:
the second pool groove is arranged at the lower end of the cover plate and is communicated with the first round groove;
The third pond groove, third pond groove sets up on the up end of base plate with second pond groove phase-match, third pond groove and second circular slot intercommunication, second pond groove and third pond groove are respectively with horizontal plane interval array three, and the size is the same, aligns perpendicularly in proper order.
As a further improvement of the invention, the temperature control zone comprises a fourth tank which is arranged on the upper end surface of the base plate and used for heating the liquid to a preset temperature range, and the fourth tank is communicated with the third tank.
As a further improvement of the present invention, the mixing zone includes:
the third round groove is arranged on the upper end face of the substrate and communicated with the upper end face of the substrate;
and the bending groove is arranged at the upper end of the substrate and communicated with the third round groove.
As a further improvement of the invention, the detection area comprises a fourth round groove, the fourth round groove is arranged on the upper end surface of the substrate and is communicated with the bending groove, and a straight groove for draining liquid is arranged on one side, away from the bending groove, of the fourth round groove on the upper end surface of the substrate.
As a further improvement of the invention, the upper end face of the base plate is bonded with the lower end face of the cover plate through an adhesive film, and the base plate and the cover plate are both made of polymethyl methacrylate.
As a further improvement of the invention, the lower end surface of the base plate is provided with a temperature control module at the lower end of the fourth tank.
The invention also provides a detection method of the microfluidic chip, which comprises the following steps:
S1, injecting a treated body fluid sample of a patient into a microfluidic chip through a first tank, and realizing the split flow into a first circular tank and a second circular tank under the action of an external power device;
S2, flowing into the second pool tank and the third pool tank under the action of power to carry out mixed reaction with the amplifying enzyme;
S3, after the mixed reaction, entering a fourth tank, and heating to a preset temperature range for amplification;
S4, after amplification, uniformly mixing the amplified product through a third circular groove and a bending groove under the drive of power;
s5, finally entering a fourth circular groove, realizing detection by a fluorescence means, and discharging the detected waste liquid through a straight groove.
As a further improvement of the invention, the external power device is a power pump, and the liquid control is from an external device.
Compared with the prior art, the invention has the beneficial effects that:
the device has the advantages of simple structure, high integration level and less required detection liquid, can better promote the stability of detection through external driving force, can realize the uniform distribution of samples in one sample inlet to different reaction chambers, realize the simultaneous amplification of multiple target partitions of one sample inlet, can prevent cross contamination between different reaction tanks, reduce the operation of multiple sample injections of operators, realize high-sensitivity, simple and stable detection, do not need professional laboratory environment, do not need professional training, and are easy and convenient to operate, thereby being suitable for home detection of ordinary masses and detection under the conditions of some field environments or lack of professionals and equipment.
Drawings
FIG. 1 is an overall exploded view of the present invention;
FIG. 2 is an overall perspective view of the present invention;
fig. 3 is a top view of a substrate of the present invention.
In the figure: 1. a substrate; 2. a cover plate; 3. a sample injection area; 31. a first tank; 32. a first circular groove; 33. a second circular groove; 4. a reaction zone; 41. a second tank; 42. a third tank; 5. a temperature control zone; 51. a fourth tank; 6. a mixing zone; 61. a third circular groove; 62. a curved slot; 7. a detection zone; 71. a fourth circular groove; 8. straight grooves.
Detailed Description
In order to make the technical problems, technical schemes and beneficial effects to be solved more clear, the invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
It will be understood that when an element is referred to as being "mounted," "connected," or "disposed" on another element, it can be directly on the other element or be indirectly on the other element. It is to be understood that the terms "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like indicate or are based on the orientation or positional relationship shown in the drawings, merely to facilitate describing the present invention and simplify the description, and do not indicate or imply that the devices or elements referred to must have the orientation specific to the specification, be constructed and operated in the specific orientation, and thus should not be construed as limiting the present invention.
As a further refinement of the present invention, the terms "first," "second," "third," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature.
Example 1
Referring to fig. 1-3, the present invention provides a technical solution: the micro-fluidic chip comprises a substrate 1 and a cover plate 2 which is arranged at the upper end of the substrate 1 in a sealing manner, wherein a micro-channel used for flowing in and out of one side of the substrate 1 from the upper end surface of the cover plate 2 is formed between the substrate 1 and the upper cover plate 2 after the substrate 1 and the upper cover plate 2 are closed, and the micro-channel comprises a sample injection area 3, a reaction area 4, a temperature control area 5, a mixing area 6 and a detection area 7.
The sample feeding area 3 comprises a first tank groove 31, a first round groove 32 and a second round groove 33, wherein the first tank groove 31 is formed in the upper end face of the cover plate 2, the first round groove 32 is formed in the lower end face of the cover plate 2 and is communicated with the first tank groove 31, the second round groove 33 is formed in the upper end of the base plate 1 and is matched with the first round groove 32, and the first round groove 32 and the second round groove 33 are respectively arranged in an array at intervals with the same size and are vertically aligned in sequence.
The reaction zone 4 comprises a second tank 41 and a third tank 42, the second tank 41 is arranged at the lower end of the cover plate 2 and communicated with the first round tank 32, the third tank 42 is arranged on the upper end face of the base plate 1 and matched with the second tank 41, the third tank 42 is communicated with the second round tank 33, the second tank 41 and the third tank 42 are respectively arranged in an array at intervals with the same size and are vertically aligned in sequence.
The temperature control area 5 comprises a fourth tank 51, the fourth tank 51 is arranged on the upper end surface of the base plate 1 and is used for heating the liquid to a preset temperature range, and the fourth tank 51 is communicated with the third tank 42.
The mixing zone 6 includes a third circular groove 61 and a curved groove 62, the third circular groove 61 is formed on the upper end surface of the base plate 1 and communicated with the curved groove 62 is formed on the upper end of the base plate 1 and communicated with the third circular groove 61.
The detection area 7 includes a fourth circular groove 71, the fourth circular groove 71 is disposed on the upper end surface of the substrate 1 and is communicated with the curved groove 62, and a straight groove 8 for draining liquid is disposed on the upper end surface of the substrate 1 at a side of the fourth circular groove 71 away from the curved groove 62.
The upper end face of the base plate 1 is bonded with the lower end face of the cover plate 2 through an adhesive film, and the base plate 1 and the cover plate 2 are both made of polymethyl methacrylate.
The lower end surface of the substrate 1 is provided with a temperature control module at the lower end of the fourth tank 51.
Example 2
The invention also provides a technical scheme that: the detection method of the microfluidic chip comprises the following steps:
s1, injecting a treated body fluid sample of a patient into a microfluidic chip through a first tank 31, and realizing the flow division into a first circular groove 32 and a second circular groove 33 under the action of an external power device;
S2, flowing into the second tank 41 and the third tank 42 under the action of power to carry out mixed reaction with the amplification enzyme;
s3, after the mixed reaction, the mixture enters a fourth tank 51 and is heated to a preset temperature range for amplification;
s4, after amplification, uniformly mixing the amplified materials through a third circular groove 61 and a bending groove 62 under the drive of power;
s5, finally entering the fourth circular groove 71, realizing detection by a fluorescence means, and discharging the detected waste liquid through the straight groove 8.
The external power device is a power pump, and the liquid control is from an external device.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. The utility model provides a micro-fluidic chip, includes base plate (1) and seals apron (2) of setting in base plate (1) upper end, its characterized in that: the micro-channel for flowing liquid from the upper end face of the cover plate (2) into the micro-channel flowing out of one side of the base plate (1) is arranged between the base plate (1) and the upper cover plate (2), and the micro-channel comprises a sample injection area (3), a reaction area (4), a temperature control area (5), a mixing area (6) and a detection area (7).
2. A microfluidic chip according to claim 1, wherein: the sample injection area (3) comprises:
The first pool groove (31) is arranged on the upper end surface of the cover plate (2);
The first round groove (32) is arranged on the lower end surface of the cover plate (2) and is communicated with the first pool groove (31);
The second round groove (33), second round groove (33) set up in the upper end of base plate (1) and first round groove (32) assorted, first round groove (32) and second round groove (33) are respectively with horizontal plane interval array three, and the size is the same, aligns perpendicularly in proper order.
3. A microfluidic chip according to claims 1 and 2, wherein: the reaction zone (4) comprises:
The second tank groove (41) is arranged at the lower end of the cover plate (2) and is communicated with the first round groove (32);
The third pond groove (42), third pond groove (42) set up on the up end of base plate (1) with second pond groove (41) phase-match, third pond groove (42) and second circular slot (33) intercommunication, second pond groove (41) and third pond groove (42) are respectively with horizontal plane interval array three, and the size is the same, vertical alignment in proper order.
4. A microfluidic chip according to claim 3, wherein: the temperature control area (5) comprises a fourth tank groove (51), the fourth tank groove (51) is arranged on the upper end face of the base plate (1) and used for heating liquid to a preset temperature range, and the fourth tank groove (51) is communicated with the third tank groove (42).
5. The microfluidic chip according to claim 4, wherein: the mixing zone (6) comprises:
A third circular groove (61), wherein the third circular groove (61) is arranged on the upper end surface of the substrate (1) and communicated with the upper end surface;
And a curved groove (62), wherein the curved groove (62) is arranged at the upper end of the substrate (1) and is communicated with the third round groove (61).
6. The microfluidic chip according to claim 5, wherein: the detection area (7) comprises a fourth round groove (71), the fourth round groove (71) is arranged on the upper end face of the base plate (1) and is communicated with the bending groove (62), and a straight groove (8) for draining liquid is arranged on one side, far away from the bending groove (62), of the fourth round groove (71) on the upper end face of the base plate (1).
7. A microfluidic chip according to claim 1, wherein: the upper end face of the base plate (1) is bonded with the lower end face of the cover plate (2) through an adhesive film, and the base plate (1) and the cover plate (2) are both made of polymethyl methacrylate.
8. The microfluidic chip according to claim 4, wherein: the lower end face of the base plate (1) is provided with a temperature control module at the lower end of the fourth tank (51).
9. The method for detecting a microfluidic chip according to claims 1 to 8, wherein: the detection method comprises the following steps:
S1, injecting a treated body fluid sample of a patient into a microfluidic chip through a first tank (31), and realizing the flow division into a first circular groove (32) and a second circular groove (33) under the action of an external power device;
S2, flowing into a second pool tank (41) and a third pool tank (42) under the action of power to carry out mixed reaction with the amplification enzyme;
s3, after the mixed reaction, the mixture enters a fourth tank (51) and is heated to a preset temperature range for amplification;
S4, after amplification, uniformly mixing the amplified product through a third round groove (61) and a bending groove (62) under the drive of power;
s5, finally entering a fourth circular groove (71) and realizing detection by a fluorescence means, and discharging the detected waste liquid through a straight groove.
10. The method for detecting a microfluidic chip according to claim 9, wherein: the external power device is a power pump, and the liquid control is from external equipment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410113889.8A CN117987250A (en) | 2024-01-27 | 2024-01-27 | Microfluidic chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410113889.8A CN117987250A (en) | 2024-01-27 | 2024-01-27 | Microfluidic chip |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117987250A true CN117987250A (en) | 2024-05-07 |
Family
ID=90890142
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410113889.8A Pending CN117987250A (en) | 2024-01-27 | 2024-01-27 | Microfluidic chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117987250A (en) |
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2024
- 2024-01-27 CN CN202410113889.8A patent/CN117987250A/en active Pending
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