Integrated micro-fluidic chip
By designing an integrated microfluidic chip, the problems of complex experimental procedures and high equipment costs in nucleic acid extraction and amplification technologies have been solved, achieving the effects of simplified operation and reduced equipment costs.
Patent Information
- Application Number
- CN202422838933.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-21
- Publication Date
- 2025-11-14
- Estimated Expiration
- 2034-11-21
AI Technical Summary
Existing nucleic acid extraction and amplification technologies suffer from problems such as complex experimental procedures, high equipment costs, large footprints, and high requirements for laboratory environment, especially fully automated equipment solutions.
Design an integrated microfluidic chip that connects the through-hole to the receiving cavity by moving a sliding cover, and uses an elastic sealing ring in the cylindrical channel to ensure airtightness, thereby realizing nucleic acid extraction, methylation purification and PCR detection.
It simplifies the experimental process, reduces equipment costs, reduces requirements for the laboratory environment, and improves experimental efficiency and equipment portability.
Smart Images
Figure CN223547981U_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of biomedical engineering, specifically to an integrated microfluidic chip. Background Technology
[0002] Nucleic acids, collectively known as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), are biological macromolecules composed of numerous nucleotide monomers and are among the most fundamental substances of life. Nucleic acids are a class of biopolymers, essential building blocks of all known life forms, and the most important of all biomolecules, widely present in all animal and plant cells and microorganisms. Nucleic acids are composed of nucleotides, and nucleotide monomers are composed of a pentose sugar, a phosphate group, and a nitrogenous base. If the pentose sugar is ribose, the resulting polymer is RNA; if the pentose sugar is deoxyribose, the resulting polymer is DNA.
[0003] Methylation refers to the catalytic transfer of methyl groups from an active methyl compound to other compounds, forming various methyl compounds or chemically modifying certain proteins or nucleic acids to form methylated products. Within biological systems, methylation is enzymatically catalyzed and involves heavy metal modification, regulation of gene expression, regulation of protein function, and ribonucleic acid processing.
[0004] In vertebrates, DNA methylation generally occurs at CpG sites (cytosine-phosphate-guanine sites, i.e., sites in the DNA sequence where cytosine is immediately followed by guanine). Cytosine is converted to 5-methylcytosine by DNA methyltransferases. Approximately 80%-90% of CpG sites in the human genome are methylated, but certain specific regions, such as CpG islands rich in cytosine and guanine, remain unmethylated. This is related to promoters, which comprise 56% of mammalian genes, including all widely expressed genes. 1%-2% of the human genome is composed of CpG sites, and CpG methylation is inversely proportional to transcriptional activity.
[0005] Polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. It can be viewed as a special form of DNA replication outside of living organisms. The most significant characteristic of PCR is its ability to dramatically increase minute amounts of DNA. Therefore, whether it's ancient fossils, the remains of historical figures, or hair, skin, or blood left behind by a murderer decades ago, as long as a tiny amount of DNA can be isolated, it can be amplified using PCR for comparison. This is the power of "trace evidence." PCR utilizes the fact that DNA denatures into single strands at 95°C in vitro. At a lower temperature (often around 60°C), primers bind to the single strands according to the principle of complementary base pairing. The temperature is then adjusted to the optimal reaction temperature of DNA polymerase (around 72°C), where the DNA polymerase synthesizes complementary strands along the phosphate-to-pentose (5'-3') direction. A PCR instrument based on polymerase is essentially a temperature control device, capable of precisely controlling the denaturation, annealing, and extension temperatures.
[0006] Currently, there are three main methods for nucleic acid extraction and amplification: 1) Manual experimental protocol: In this protocol, researchers use pipettes, constant temperature shakers, mixers, and other equipment in a specific experimental environment, following the reagent instructions, to extract DNA from samples, methylate and purify it, and finally perform PCR amplification. However, this protocol is complex, time-consuming, and inefficient; it requires a wide variety of small devices and places extremely high demands on the laboratory environment. 2) Semi-automatic equipment experimental protocol: In this protocol, researchers use pipettes to add reagents to the equipment consumables according to the equipment's instructions, and then run the semi-automatic equipment for nucleic acid extraction and methylation conversion experiments. After the experiment, the researcher manually transfers the DNA for PCR amplification. However, this protocol requires manual monitoring and sample addition during the experiment and places extremely high demands on the laboratory environment. 3) Fully automated equipment experimental protocol: Researchers directly operate automated equipment, which automatically completes nucleic acid extraction, methylation conversion, and PCR amplification. This protocol requires large equipment footprint, is expensive, and is difficult to develop. Utility Model Content
[0007] To address the aforementioned deficiencies, this application provides an integrated microfluidic chip. By moving the sliding cover, the first through hole is connected to different cavities, thereby enabling experiments (e.g., DNA extraction, methylation purification, and PCR detection). Because an elastic sealing ring is provided inside the cylindrical channel, the cavities are not connected to the outside during the experiment, thus ensuring the chip's airtightness.
[0008] This application provides the following technical solution.
[0009] This application provides an integrated microfluidic chip, wherein the chip includes a chip body and a sliding cover, the sliding cover being movably disposed on the top of the chip body, and the sliding cover being movable along the extension direction of the chip body.
[0010] The chip body has at least two recessed cavities along its extension direction, and each cavity is independent of the others.
[0011] At least two first through holes are provided on the sliding cover, and a cylindrical channel protruding from the surface of the sliding cover is provided on the side of the sliding cover away from the chip body, and the cylindrical channel is located above the first through holes. An elastic sealing ring is provided in the cylindrical channel.
[0012] Furthermore, at least two of the first through holes are arranged sequentially along the extension direction of the sliding cover.
[0013] Furthermore, a second through hole is provided on the sliding cover, and an opening channel protruding from the surface of the sliding cover is provided on the side surface of the sliding cover opposite to the chip body, and the opening channel is located above the second through hole.
[0014] Furthermore, a channel cover for closing the opening channel is provided at the top of the opening channel.
[0015] Furthermore, the opening channel and the cylindrical channel are on the centerline of the side surface of the sliding cover opposite to the chip body;
[0016] The height of the open channel is less than the height of the cylindrical channel.
[0017] Furthermore, a mounting plate perpendicular to the surface of the sliding cover is provided on the side of the sliding cover away from the chip body. The mounting plate is located between the opening channel and the cylindrical channel, and one end of the mounting plate is connected to the outer wall of the cylindrical channel.
[0018] Furthermore, the sliding cover has a groove on the side near the chip body, and the chip body has a sliding piece that cooperates with the groove on the side near the sliding cover.
[0019] The slide has an opening that mates with the receiving cavity.
[0020] Furthermore, the plurality of receiving cavities are divided into a first receiving cavity, a second receiving cavity, and a third receiving cavity.
[0021] The first receiving cavity is located near the end of the chip body. The first receiving cavity includes a first receiving cavity body and a detection tube detachably connected to its bottom, and the first receiving cavity body is in communication with the detection tube.
[0022] The opening size of the second receiving cavity is smaller than the opening size of the third receiving cavity.
[0023] Furthermore, the second receiving cavity includes a second receiving cavity A and a second receiving cavity B, wherein the second receiving cavity A is used to store reagents and the second receiving cavity B is used to provide experimental space for experiments;
[0024] The third accommodating cavity includes a third accommodating cavity A, a third accommodating cavity B, and a third accommodating cavity C. The third accommodating cavity A is used to store reagents and also provides experimental space; the third accommodating cavity B is used to provide experimental space; and the third accommodating cavity C is used to store reagents or instruments.
[0025] Furthermore, the number of the first receiving cavity is at least one, the number of the second receiving cavity is at least two, and the number of the third receiving cavity is at least three.
[0026] Furthermore, the openings of the first receiving cavity, the second receiving cavity, and the third receiving cavity are circular, elliptical, or n-sided with n greater than or equal to 3, respectively.
[0027] The integrated microfluidic chip provided in this application allows for experiments (such as DNA extraction, methylation purification, and PCR detection) to be conducted by moving the sliding cover and connecting the first through hole with different cavities through the sliding cover. Because the cylindrical channel is equipped with an elastic sealing ring, the cavities are not connected to the outside during the experiment, thus ensuring the chip's airtightness. Attached Figure Description
[0028] Figure 1 This is a schematic diagram of the integrated microfluidic chip structure provided in this application.
[0029] Figure 2 This is a schematic diagram of the sliding cover provided in this application.
[0030] Explanation of reference numerals in the attached figures
[0031] 1-Sliding cover, 2-Chip body, 3-Cylindrical channel, 4-Open channel, 5-Channel cover, 6-Mounting plate, 7-First receiving cavity, 8-Sliding groove, 9-Sliding piece, 10-Detection tube, 11-Third receiving cavity, 12-Second receiving cavity, 13-Injector, 14-Magnetic rod sleeve. Detailed Implementation
[0032] The following description provides exemplary embodiments of this application, including various details to aid understanding, and should be considered merely exemplary. Therefore, those skilled in the art will recognize that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of this application. Similarly, for clarity and brevity, descriptions of well-known functions and structures are omitted in the following description.
[0033] like Figure 1 as well as Figure 2 As shown, this application provides an integrated microfluidic chip, wherein the chip includes a chip body 2 and a sliding cover 1. The sliding cover 1 is movably disposed on the top of the chip body 2, and in use, the sliding cover 1 moves along the extending direction of the chip body 2.
[0034] The chip body 2 has at least two recessed cavities along its extending direction, and each cavity is independent and not connected to the others.
[0035] At least two first through holes are provided on the sliding cover 1, and a cylindrical channel 3 protruding from the surface of the sliding cover 1 is provided on the side surface of the sliding cover 1 away from the chip body 2. The cylindrical channel 3 is located above the first through holes. An elastic sealing ring is provided in the cylindrical channel 3. In use, the first through hole communicates with the receiving cavity.
[0036] In this document, the extending direction of the chip body 2 is the long side direction of the chip, and the extending direction of the chip body 2 is perpendicular to the recessed direction of the receiving cavity. The extending direction of the chip body 2 is consistent with the extending direction of the sliding cover 1.
[0037] The number of first through holes on the sliding cover 1 can be 2, 3, 4, 5, 6, 7, etc., and the number of first through holes can be determined according to actual needs.
[0038] The inner diameter of the first through-hole is 1mm-100mm, for example, it can be 1mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, etc., and the spacing is 10mm-30mm, for example, it can be 10mm, 15mm, 20mm, 25mm, 30mm, etc. The first through-hole can be used to mate with a disposable syringe or a magnetic rod sleeve to seal or experiment on the chip body. The inner diameters of multiple first through-holes can be the same or different. For example, the inner diameter of the first through-hole used to fit with a disposable syringe is smaller than the inner diameter of the first through-hole used to fit with a magnetic rod sleeve.
[0039] Furthermore, at least two of the first through holes are arranged sequentially along the extending direction of the sliding cover 1. In some embodiments, all the first through holes are arranged sequentially along the extending direction of the sliding cover 1, and adjacent first through holes are equally spaced. This spacing can be matched with the receiving cavity of the chip body, and the spacing is consistent with the distance between the receiving cavity, making it easy for the chip to be adapted to the device.
[0040] Furthermore, the number of the cylindrical channels 3 is equal to the number of the first through holes, and the inner diameter of the cylindrical channels 3 is greater than or equal to the inner diameter of the first through holes. The bottom edge of the cylindrical channels 3 is arranged around the edge of the first through holes.
[0041] The inner diameter of the cylindrical channel 3 is 2mm-120mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, 110mm, 120mm, etc., and its height is 5mm-120mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, 110mm, 120mm, etc. This design facilitates the placement and removal of disposable syringes and magnetic rod sheaths.
[0042] In some embodiments, the sliding cover 1 is provided with three first through holes penetrating the surface of the sliding cover 1, and the three first through holes are arranged sequentially along the extending direction of the sliding cover 1. A cylindrical channel 3 is provided around the edge of the first through holes on the surface of the sliding cover 1, and the cylindrical channel 3 corresponds one-to-one with the first through holes.
[0043] Furthermore, the outer periphery of the sealing ring is connected to the inner wall of the cylindrical channel 3, and the sealing ring has an opening in the middle. The sealing ring is an elastic sealing ring. In use, the syringe 13 or the magnetic rod sleeve 14 passes through the opening and enters the first through hole, thus entering the receiving cavity. The inner diameter of the opening of the sealing ring is smaller than the outer diameter of the syringe 13 and the magnetic rod sleeve 14. This design isolates the receiving cavity from the external environment, thereby ensuring the chip's sealing performance.
[0044] During the methylation process of the chip, the syringe 13 or the magnetic rod sleeve 14 extends into the receiving cavity through the cylindrical channel 3. The sealing ring inside the cylindrical channel 3 is in tight contact with the side wall of the syringe 13 or the magnetic rod sleeve 14 to prevent communication between the outside and the receiving cavity. When the sliding cover 1 moves along the extension direction of the chip body 2, the syringe 13 or the magnetic rod sleeve 14 moves out of the receiving cavity, but the syringe 13 or the magnetic rod sleeve 14 does not extend out of the cylindrical channel 3. That is, the end of the syringe 13 or the magnetic rod sleeve 14 near the receiving cavity is still between the sealing ring and the receiving cavity to ensure that the chip body 2 is not connected to the external environment.
[0045] When the chip is used for DNA extraction and amplification, some of the multiple cavities contain experimental reagents, some provide operating space for DNA extraction, and some provide space for DNA amplification.
[0046] In this application, a second through hole is provided on the sliding cover 1, and an opening channel 4 protruding from the surface of the sliding cover 1 is provided on the side surface of the sliding cover 1 opposite to the chip body 2, and the opening channel 4 is located directly above the second through hole.
[0047] The number of the second through holes can be 1, 2, 3, 4, 5, 6, 7, 8, 9, etc., and the number of the second through holes can be determined according to actual needs.
[0048] The inner diameter of the second through hole is 2mm-100mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, etc. The inner diameter of the opening channel 4 is 5mm-120mm, for example, it can be 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, 110mm, 120mm, etc., and the height is 2mm-100mm, for example, it can be 2mm, 5mm, 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm, 50mm, 55mm, 60mm, 65mm, 70mm, 75mm, 80mm, 85mm, 90mm, 95mm, 100mm, etc. The experimental sample is added into the corresponding receiving cavity through the second through hole and the opening channel 4.
[0049] Furthermore, the number of second through holes is the same as the number of opening channels 4. On the surface of the sliding cover 1, the bottom of the opening channel 4 is arranged around the second through hole, and the inner diameter of the opening channel 4 is greater than or equal to the inner diameter of the second through hole. The center of the second through hole is located on the center line of all the first through holes.
[0050] Furthermore, a channel cover 5 for closing the opening channel 4 is provided at the top of the opening channel 4. In some embodiments, the opening channel 4 and the channel cover 5 are connected by a hinge.
[0051] The opening channel 4 and the cylindrical channel 3 are on the center line of the side surface of the sliding cover 1 opposite to the chip body 2.
[0052] The height of the open channel 4 is less than the height of the cylindrical channel 3.
[0053] In this application, a mounting plate 6 perpendicular to the surface of the sliding cover 1 is provided on the side surface of the sliding cover 1 away from the chip body 2. The mounting plate 6 is located between the opening channel 4 and the cylindrical channel 3, and one end of the mounting plate 6 is connected to the outer wall of the cylindrical channel 3.
[0054] The mounting plate 6 is an arc-shaped plate with right-angled sides. It is used in conjunction with the equipment to move the sliding cover 1, making it easier for the experimenter to grasp the chip.
[0055] The lengths of the right-angled sides of the mounting plate 6 are L1 and L2, respectively. L1 is the side length that contacts the sliding cover 1, and L2 is the side length that contacts the cylindrical channel 3.
[0056] In this application, the sliding cover 1 has a sliding groove 8 on the side near the chip body 2, and the chip body 2 has a sliding piece 9 on the side near the sliding cover 1 that cooperates with the sliding groove 8. An opening that cooperates with the receiving cavity is provided on the sliding piece 9.
[0057] The extension direction of the slide groove 8, the extension direction of the slide cover 1, and the extension direction of the slide plate 9 are consistent.
[0058] The chip body 2 is composed of a slider 9 and a receiving cavity body. The slider 9 is located on the top of the receiving cavity body and is integrally formed with the receiving cavity body. The width of the slider 9 is greater than the width of the receiving cavity body, and the opening on the slider 9 communicates with the receiving cavity body.
[0059] When the sliding cover 1 needs to move, the sliding groove 8 moves along the extending direction of the sliding piece 9, thereby making the first through hole communicate with the corresponding receiving cavity.
[0060] In this application, the plurality of receiving cavities are divided into a first receiving cavity 7, a second receiving cavity 12, and a third receiving cavity 11. The first receiving cavity 7 is located near the end of the chip body 2. The first receiving cavity 7 includes a first receiving cavity body and a detection tube 10 detachably connected to its bottom, and the first receiving cavity body communicates with the detection tube 10. The purpose of designing the first receiving cavity 7 in this way is to ensure that the tip of the disposable pipette and the internal reagent are in a sealed state during the process of transferring liquid to the detection tube 10.
[0061] The opening size of the second receiving cavity 12 is smaller than the opening size of the third receiving cavity 11.
[0062] In this article, the opening size refers to the diameter or equivalent diameter.
[0063] The capacity of the first receiving cavity 7 is 1000ml-5000ml, for example, it can be 1000ml, 1500ml, 2000ml, 2500ml, 3000ml, 3500ml, 4000ml, 4500ml, 5000ml, etc.
[0064] The capacity of the second receiving cavity 12 is 100ml-3000ml, for example, it can be 100ml, 300ml, 500ml, 800ml, 1000ml, 1500ml, 2000ml, 2500ml, 3000ml, etc.
[0065] The capacity of the third receiving cavity 11 is 100ml-15000ml, for example, it can be 100ml, 300ml, 500ml, 800ml, 1000ml, 1500ml, 2000ml, 2500ml, 3000ml, 4000ml, 5000ml, 8000ml, 10000ml, 12000ml, 15000ml, etc.
[0066] The number of the first receiving cavity 7 is at least 1, the number of the second receiving cavity 12 is at least 2, and the number of the third receiving cavity 11 is at least 3.
[0067] The number of the first receiving cavities 7 can be 1, 2, 3, 4, 5, 6, 7, 8, etc., the number of the second receiving cavities 12 can be 2, 3, 4, 5, 6, 7, 8, etc., and the number of the third receiving cavities 11 can be 3, 4, 5, 6, 7, 8, etc. The specific numbers can be determined according to actual needs.
[0068] Furthermore, the second receiving cavity includes a second receiving cavity A and a second receiving cavity B, wherein the second receiving cavity A is used to store reagents and the second receiving cavity B is used to provide experimental space for experiments;
[0069] The third accommodating cavity includes a third accommodating cavity A, a third accommodating cavity B, and a third accommodating cavity C. The third accommodating cavity A is used to store reagents and also provides experimental space; the third accommodating cavity B is used to provide experimental space; and the third accommodating cavity C is used to store reagents or instruments.
[0070] In some embodiments, when the chip is used for DNA extraction, methylation purification, and PCR detection, the second containment chamber includes a second containment chamber A and a second containment chamber B. The second containment chamber A stores reagents required for the DNA extraction or amplification process, and the second containment chamber B provides experimental space for the DNA extraction or amplification process. The third containment chamber includes a third containment chamber A, a third containment chamber B, and a third containment chamber C. The third containment chamber A stores reagents required for the DNA extraction process and also provides experimental space for the DNA extraction or amplification process; the third containment chamber B is used for sulfite conversion of DNA; and the third containment chamber C stores reagents or magnetic beads used in the DNA extraction process.
[0071] In some embodiments, the number of first receiving cavities 7 is one, the number of second receiving cavities 12 can be five, namely second receiving cavity A1, second receiving cavity B2, second receiving cavity A3, second receiving cavity A4, and second receiving cavity B5, and the number of third receiving cavities 11 can be eight, namely third receiving cavity A1, third receiving cavity A2, third receiving cavity B3, third receiving cavity A4, third receiving cavity A5, third receiving cavity C6, third receiving cavity C7, and third receiving cavity C8. Starting from one end of the chip body 2, the three cavities are sequentially arranged as follows: first receiving cavity 7, second receiving cavity A1, second receiving cavity B2, second receiving cavity A3, third receiving cavity A1, second receiving cavity A4, second receiving cavity B5, third receiving cavity A2, third receiving cavity B3, third receiving cavity A4, third receiving cavity A5, third receiving cavity C6, third receiving cavity C7, and third receiving cavity C8.
[0072] The first accommodating cavity 7 is used to store PCR reaction reagents and to carry out PCR reactions.
[0073] The second cavities A1, A3, and A4 are used to store reagents required during the DNA extraction or amplification process, such as paraffin oil, sulfidation reagents, and elution buffer.
[0074] The second accommodating cavities B2 and B5 are used to provide space for the DNA extraction or amplification process, and can also be used to place the syringe 13 used for injection.
[0075] The third cavities A1, A2, A4, and A5 are used to store reagents required in the DNA extraction process, such as elution buffer, rinsing buffer, and lysis buffer + magnetic beads. They also provide experimental space for DNA extraction or amplification. Additionally, the third cavity A1 can be used to place the magnetic rod sleeve 14.
[0076] The third cavity B3 is used for sulfite conversion and elution of DNA.
[0077] The third cavities C6, C7, and C8 are used to store the lysis buffer + magnetic beads, binding buffer, and washing buffer used in the DNA extraction process.
[0078] The first receiving cavity has a capacity of 500ml-4000ml, and the detection tube is a tapered tube with a capacity of 500ml-1000ml.
[0079] The capacity of the second receiving cavity A1 is 500ml-1500ml.
[0080] The capacity of the second receiving cavity B2 is 800ml-2000ml.
[0081] The capacity of the second receiving cavity A3 is 600ml-1800ml.
[0082] The capacity of the second receiving cavity A4 is 100ml-500ml.
[0083] The capacity of the second receiving cavity B5 is 800ml-2000ml.
[0084] The capacity of the third receiving cavity A1 is 3000ml-8000ml.
[0085] The capacity of the third receiving chamber A2 is 3000ml-8000ml.
[0086] The capacity of the third receiving chamber B3 is 2000ml-6000ml.
[0087] The capacity of the third receiving cavity A4 is 3000ml-8000ml.
[0088] The capacity of the third receiving cavity A5 is 5000ml-15000ml.
[0089] The capacity of the third receiving cavity C6 is 1000ml-5000ml.
[0090] The capacity of the third receiving chamber C7 is 3000ml-8000ml.
[0091] The capacity of the third receiving chamber C8 is 100ml-500ml.
[0092] The openings of the first receiving cavity 7, the second receiving cavity, and the third receiving cavity 11 are circular, elliptical, or n-sided, with n being greater than or equal to 3. The n-sided shape can be rectangular, regular hexagonal, regular octagonal, regular dodecagonal, regular 24-sided, etc.
[0093] In some embodiments, the opening of the first receiving cavity 7 is circular, the opening of the second receiving cavity 12 is circular, and the opening of the third receiving cavity 11 is circular or rectangular.
[0094] Example
[0095] The materials and test methods used in the embodiments of this application are described in a general and / or specific manner. In the following embodiments, unless otherwise specified, % means wt%, i.e., weight percentage. Reagents or instruments used, unless otherwise specified, are all commercially available conventional reagent products.
[0096] Example 1
[0097] The integrated microfluidic chip in this embodiment is used for DNA extraction, methylation purification, and PCR detection.
[0098] The integrated microfluidic chip includes a chip body 2 and a sliding cover 1, wherein the sliding cover 1 is movably disposed on the top of the chip body 2.
[0099] The chip body 2 is provided with a first receiving cavity 7, a second receiving cavity 12, and a third receiving cavity 11 recessed into the chip body 2 along its extension direction, and each receiving cavity is independent and not connected to each other. There is one first receiving cavity 7, and there can be five second receiving cavities 12, namely second receiving cavity A1, second receiving cavity B2, second receiving cavity A3, second receiving cavity A4, and second receiving cavity B5. There can be eight third receiving cavities 11, namely third receiving cavity A1, third receiving cavity A2, third receiving cavity B3, third receiving cavity A4, third receiving cavity A5, third receiving cavity C6, third receiving cavity C7, and third receiving cavity C8. Starting from one end of the chip body 2, there are sequentially arranged first receiving cavity 7, second receiving cavity A1, second receiving cavity B2, second receiving cavity A3, third receiving cavity A1, second receiving cavity A4, second receiving cavity B5, third receiving cavity A2, third receiving cavity B3, third receiving cavity A4, third receiving cavity A5, third receiving cavity C6, third receiving cavity C7, and third receiving cavity C8. The opening of the first receiving cavity 7 is circular, the openings of the second receiving cavities are all circular, the openings of the third receiving cavities A1 and C8 are both circular, and the openings of the third receiving cavities A2, B3, A4, A5, C6, and C7 are all rectangular. The first receiving cavity 7 consists of a main body and a detection tube 10 detachably connected to its bottom, and the main body of the first receiving cavity 7 is in communication with the detection tube 10.
[0100] The first receiving cavity has a capacity of 2000ml, and the detection tube is a tapered tube with a capacity of 900ml.
[0101] The capacity of the second receiving cavity A1 is 950 ml.
[0102] The capacity of the second receiving cavity B2 is 1500ml.
[0103] The capacity of the second receiving cavity A3 is 950 ml.
[0104] The capacity of the second receiving cavity A4 is 400ml.
[0105] The capacity of the second receiving cavity B5 is 1500ml.
[0106] The capacity of the third receiving cavity A1 is 5000ml.
[0107] The third receiving chamber A2 has a capacity of 5000 ml.
[0108] The third receiving chamber B3 has a capacity of 3500ml.
[0109] The third receiving chamber A4 has a capacity of 5000ml.
[0110] The third receiving chamber A5 has a capacity of 9000 ml.
[0111] The third receiving chamber C6 has a capacity of 2000 ml.
[0112] The third receiving chamber C7 has a capacity of 5000ml.
[0113] The third receiving chamber C8 has a capacity of 300 ml.
[0114] The sliding cover 1 has three first through holes and one second through hole, arranged sequentially along the extension direction of the sliding cover 1. The three first through holes are evenly spaced at 15.5 mm intervals. A cylindrical channel 3 is provided around the edge of the first through hole, and an open channel 4 is provided around the edge of the second through hole on the surface of the sliding cover 1. An elastic sealing ring is provided around the inner wall of the cylindrical channel 3, with an opening in the center. A channel cover 5 for closing the open channel 4 is hinged to the top of the open channel 4. The height of the open channel 4 is 4.5 mm, and its inner diameter is 12 mm. The height of the cylindrical channel 3 is 5.5 mm, and its inner diameter is 11 mm. The inner diameter of the first through hole is 8 mm, and the inner diameter of the second through hole is 11 mm.
[0115] An arc-shaped mounting plate 6 with a right-angled edge is provided on the surface of the sliding cover 1 opposite to the chip body 2. The mounting plate 6 is located between the opening channel 4 and the cylindrical channel 3, and one end of the mounting plate 6 is connected to the outer wall of the cylindrical channel 3. The sliding cover 1 has a sliding groove 8 on the side near the chip body 2, and the chip body 2 has a sliding piece 9 on the side near the sliding cover 1 that cooperates with the sliding groove 8. An opening that cooperates with the receiving cavity is provided on the sliding piece 9.
[0116] In this embodiment, the chip is used by first moving the sliding cover 1 to move the opening channel 4 directly above the third receiving cavity b5, and adding the plasma sample into the third receiving cavity b5 through the opening channel 4; then, moving the sliding cover 1 so that the receiving cavity containing the syringe 13 is directly above the receiving cavity containing the reagents needed for DNA lysis, drawing up the reagents, and then moving the syringe 13 directly above the third receiving cavity b5 through the sliding cover 1, thereby transferring the reagents needed for DNA lysis into the third receiving cavity b5; then, moving the receiving cavity containing the magnetic rod sleeve 14 directly above the third receiving cavity b5, and performing a mixing and oscillating action within the third receiving cavity b5 through the magnetic rod sleeve 14, thereby lysing the DNA from the plasma; after the lysis process is completed, the sample is removed by moving the sliding cover 1 to the receiving cavity. The sliding cover 1 transfers the magnetic beads to the third receiving chamber b4 and elutes by shaking. After elution, the sliding cover 1 is moved to transfer the magnetic beads to the third receiving chamber b3, and the reagents required for sulfite conversion are also transferred to the third receiving chamber b3. Sulfite conversion is performed in the third receiving chamber b3, and the beads are eluted by shaking. After elution, the sliding cover 1 is moved to transfer the magnetic beads to the third receiving chamber b2 through the magnetic rod sleeve 14 for washing. After washing, the sliding cover 1 is moved to transfer the magnetic beads to the third receiving chamber b1 for elution by shaking. After elution, the sliding cover 1 is moved to transfer the magnetic beads to the third receiving chamber b2, and the eluent is transferred to the first receiving chamber 7. Then, the sliding cover 1 is moved to transfer the paraffin oil in the second receiving chamber a1 to the first receiving chamber 7 for PCR reaction. Therefore, the chip of this application uses two disposable syringes 13 to transfer liquid; a magnetic rod is used to shake and mix the liquid to wash off the magnetic beads; all reagents required for the experiment are pre-packaged in the detection box; the same reagent is stored in the same well and transferred by disposable syringe 13 when used; the detection tube 10 for PCR amplification is independently designed and can be used in a conventional PCR instrument. The structure is simple, the operation is convenient, and the equipment cost is greatly reduced.
[0117] Although the embodiments of this application have been described above in conjunction with the accompanying drawings, this application is not limited to the specific embodiments and application fields described above. The specific embodiments described above are merely illustrative and instructive, not restrictive. Those skilled in the art can make many other forms based on the guidance of this specification and without departing from the scope of protection of the claims of this application, and these are all within the scope of protection of this application.
Claims
1. An integrated microfluidic chip, characterized in that, The chip includes a chip body and a sliding cover. The sliding cover is movably disposed on the top of the chip body and is capable of moving along the extending direction of the chip body. The chip body has at least two recessed cavities along its extension direction, and each cavity is independent of the others. At least two first through holes are provided on the sliding cover, and a cylindrical channel protruding from the surface of the sliding cover is provided on the side of the sliding cover away from the chip body, and the cylindrical channel is located above the first through holes. An elastic sealing ring is provided in the cylindrical channel.
2. The chip according to claim 1, characterized in that, At least two of the first through holes are arranged sequentially along the extension direction of the sliding cover.
3. The chip according to claim 1, characterized in that, A second through hole is provided on the sliding cover, and an opening channel protruding from the surface of the sliding cover is provided on the side surface of the sliding cover opposite to the chip body, and the opening channel is located above the second through hole.
4. The chip according to claim 3, characterized in that, A channel cover for closing the opening channel is provided at the top of the opening channel.
5. The chip according to claim 3, characterized in that, The opening channel and the cylindrical channel are on the center line of the side surface of the sliding cover opposite to the chip body; The height of the open channel is less than the height of the cylindrical channel.
6. The chip according to claim 3, characterized in that, A mounting plate perpendicular to the surface of the sliding cover is provided on the side of the sliding cover away from the chip body. The mounting plate is located between the opening channel and the cylindrical channel, and one end of the mounting plate is connected to the outer wall of the cylindrical channel.
7. The chip according to claim 3, characterized in that, The sliding cover has a groove on the side near the chip body, and the chip body has a sliding piece that mates with the groove on the side near the sliding cover. The slide has an opening that mates with the receiving cavity.
8. The chip according to claim 3, characterized in that, The plurality of receiving cavities are divided into a first receiving cavity, a second receiving cavity, and a third receiving cavity. The first receiving cavity is located near the end of the chip body. The first receiving cavity includes a first receiving cavity body and a detection tube detachably connected to its bottom, and the first receiving cavity body is in communication with the detection tube. The opening size of the second receiving cavity is smaller than the opening size of the third receiving cavity.
9. The chip according to claim 8, characterized in that, The second containment cavity includes a second containment cavity A and a second containment cavity B. The second containment cavity A is used to store reagents, and the second containment cavity B is used to provide experimental space for experiments. The third accommodating cavity includes a third accommodating cavity A, a third accommodating cavity B, and a third accommodating cavity C. The third accommodating cavity A is used to store reagents and also provides experimental space; the third accommodating cavity B is used to provide experimental space; and the third accommodating cavity C is used to store reagents or instruments.
10. The chip according to claim 8, characterized in that, The number of the first receiving cavity is at least one, the number of the second receiving cavity is at least two, and the number of the third receiving cavity is at least three; or The openings of the first receiving cavity, the second receiving cavity, and the third receiving cavity are circular, elliptical, or n-sided, respectively, where n is greater than or equal to 3.