CN219653039U - Miniature enzyme thermal biosensor based on thermoelectric self-energy supply - Google Patents
Miniature enzyme thermal biosensor based on thermoelectric self-energy supply Download PDFInfo
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- CN219653039U CN219653039U CN202320842658.1U CN202320842658U CN219653039U CN 219653039 U CN219653039 U CN 219653039U CN 202320842658 U CN202320842658 U CN 202320842658U CN 219653039 U CN219653039 U CN 219653039U
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- 108090000790 Enzymes Proteins 0.000 title claims abstract description 20
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 20
- 239000007788 liquid Substances 0.000 claims abstract description 61
- 238000001514 detection method Methods 0.000 claims abstract description 42
- 238000007789 sealing Methods 0.000 claims abstract description 32
- 239000000872 buffer Substances 0.000 claims abstract description 14
- 230000009144 enzymatic modification Effects 0.000 claims abstract description 9
- 238000006011 modification reaction Methods 0.000 claims abstract description 9
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 6
- 239000010931 gold Substances 0.000 claims description 6
- 229910052737 gold Inorganic materials 0.000 claims description 6
- 229920001609 Poly(3,4-ethylenedioxythiophene) Polymers 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 4
- 239000002184 metal Substances 0.000 claims description 4
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 4
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 2
- 229910052782 aluminium Inorganic materials 0.000 claims description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 2
- 229910052802 copper Inorganic materials 0.000 claims description 2
- 239000010949 copper Substances 0.000 claims description 2
- 229910052697 platinum Inorganic materials 0.000 claims description 2
- 238000004544 sputter deposition Methods 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 230000004044 response Effects 0.000 abstract description 2
- 229940088598 enzyme Drugs 0.000 description 20
- 239000007853 buffer solution Substances 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 238000001755 magnetron sputter deposition Methods 0.000 description 4
- 238000012544 monitoring process Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 230000007613 environmental effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 108090000204 Dipeptidase 1 Proteins 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 108010093096 Immobilized Enzymes Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 102000006635 beta-lactamase Human genes 0.000 description 1
- 239000006177 biological buffer Substances 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- -1 saliva Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model discloses a micro enzyme thermal biosensor based on thermoelectric self-energy supply, and belongs to the field of biochemical detection. The micro enzyme thermal biosensor includes: the channel layer comprises a groove embedded with the cover, a conductive circuit, a liquid flow detection channel, a signal output electrode, a cold end (p end) of the thermoelectric element, a hot end (n end) of the thermoelectric element, an enzyme modification reaction layer and a sample outlet; the cover layer is arranged above the channel layer and comprises a liquid inlet channel, a buffer liquid inlet channel, a bulge embedded with the channel layer and an accessory structure thereof; the sealing plug is arranged below the channel layer and comprises a sealing ring, an annular sealing layer and a handle layer. The utility model has the advantages of convenient carrying, simple operation, high sensitivity and quick response speed, and simultaneously has simple manufacturing process and easy mass production, and combines thermoelectric self-energy supply with the micro detection kit to realize rapid detection of enzyme thermal reaction.
Description
Technical Field
The utility model relates to the field of biochemical detection, in particular to a micro enzyme thermal biosensor based on thermoelectric self-energy supply.
Background
The biosensor uses the bio-sensitive molecules as core elements, can realize the monitoring of a plurality of important biomarkers, and can be widely applied to various aspects of medical treatment, food production, environmental monitoring and the like. The development trend of the biosensor aiming at miniaturization, miniaturized processing and the introduction of novel materials all show great potential in the aspects of accurate medical treatment, real-time monitoring and the like. Meanwhile, the development and utilization of the energy source in an actual scene are limited by the improvement of cost and continuous dependence on the energy source caused by the fine processing.
The existing enzyme thermal sensor can be used as a typical biological sensor for disease monitoring, health and medical evaluation, and can be used for evaluating biological liquids such as saliva, blood, sweat and the like. However, the method has the defects of low detection precision, continuous energy supply to maintain a constant temperature environment, difficulty in convenience and miniaturization. Therefore, the utility model combines the novel thermoelectric material Ag2Se capable of self-supplying with the enzyme heat sensor, and realizes the rapid detection of the thermal effect of the enzymatic reaction through the conversion of heat energy and electric signals.
The miniature detection kit has the advantages of less sample consumption, high response speed, multifunction integration, small volume, portability and the like, and has great application potential in the field of biological detection. However, the existing microcalorimeters lack a good combination mode with enzyme thermal reaction detection, and have the problems of low sensitivity, large occupied area, high preparation cost and the like. Therefore, there is an urgent need for a sensor that can be miniaturized, portable, sensitive, and trace-detected.
Disclosure of Invention
The first aim of the utility model is to provide a micro enzyme thermal biosensor based on thermoelectric self-energy, which solves the problems of complex manufacturing process, high manufacturing cost, large consumption of measured samples and insensitive reaction in the existing equipment.
In order to achieve the aim, the micro enzyme thermal biosensor based on thermoelectric self-energy comprises a channel layer (1), a cover layer (2) arranged above the channel layer and a sealing plug (3) arranged below the channel layer.
Furthermore, the center of the channel layer is a groove-shaped liquid flow detection channel (13), two side walls of the liquid flow detection channel are enzyme modification reaction layers (17), the top wall of the liquid flow detection channel is covered by a cover layer (2), and the liquid flow detection channel is communicated with the outside only through a liquid inlet micro-channel (21) and a buffer liquid inlet micro-channel (22) after the assembly is completed.
Further, the liquid flow detection channel (13) has a depth of 3mm, a length of 30mm and a width of 2mm.
Furthermore, the enzyme modification reaction layer is a magnetron sputtering gold film, the thickness of the gold film is 2-3 nm, and enzyme is selected from catalase, glucose oxidase, beta-lactamase, urease and the like through Au-S bond immobilized enzyme.
Further, in order to facilitate magnetron sputtering of the side wall, the junction of the bottom of the liquid flow detection channel of the channel layer and the side wall is detachable and connected with the inwardly extending groove (110) of the side wall through the bottom bulge (19).
Further, the thermoelectric elements are all located on the upper surface of the channel layer, form an angle of 90 degrees with the enzyme modification reaction layer, and are connected in series with 12-20 pairs altogether, and the cold end, namely the p end (15), of each thermoelectric element is connected with the hot end, namely the n end (16), of each thermoelectric element through a signal output electrode (14), the signal output electrodes (14) are connected through conducting circuits (12), and the detection is carried out through the external voltmeter of the conducting circuits (12).
Further, the cold end (p-end) (15) of the thermoelectric element is polyethylene dioxythiophene (PEDOT), and the hot end, i.e. the n-end (16), of the thermoelectric element is an Ag2Se film.
Further, the signal output electrode (14) is a magnetron sputtering gold electrode, and the thickness of the sputtering gold film is 30nm; the conductive circuit is made of two identical metal thin wires, and the material of the metal thin wires is platinum wires or copper wires or aluminum wires.
Further, the channel layer is connected with the cover layer (2) through a groove (11) embedded with the cover, and the sample outlet (18) is connected with the sealing plug (3) for sealing.
Further, the cover layer comprises a liquid inlet micro-channel (21), a buffer liquid inlet micro-channel (22), a bulge (23) embedded with the channel layer, a negative electrode lead outlet (25) and a positive electrode lead outlet (27).
Further, the buffer solution can be biological buffer solution or zwitterionic buffer solution, including Tris, HEPES, caps buffer solution.
Further, the liquid inlet channel (21) and the buffer liquid inlet channel (22) are horn-shaped inlets (28) which are tightly connected with the semicylindrical liquid channel (24) and the semicylindrical buffer liquid channel (26) below into a whole, wherein the diameter of the upper part of the horn-shaped inlets is 4mm, the diameter of the opening of the lower part is 1mm, the height of the semicylindrical liquid channel is 2mm, and the horn-shaped inlets are positioned in the center of the covering layer and can be completely embedded into the liquid flow detection channel below.
Further, the sealing plug (3) comprises a sealing ring (31), an annular sealing layer (32) and a handle layer (33). The sealing ring (31) is tightly attached to the annular sealing layer (32) and the sample outlet (18) of the channel layer, so that liquid can be prevented from flowing out or gas can be prevented from entering.
Further, the sealing plug is a rubber plug.
Compared with the prior art, the utility model has the beneficial effects that:
1. the utility model has the advantages of convenient carrying, simple operation, high sensitivity, low cost, high detection speed and low sample consumption, and provides a scientific and efficient detection mode for the detection of the enzyme thermal reaction.
2. According to the utility model, the micro enzyme thermal biosensor is adopted for detection, and the detection can be carried out only by a small sample size, and the thermoelectric self-energy supply and the micro reagent detection kit are combined, so that the rapid and efficient detection of environmental protection and energy self-supply is realized. Meanwhile, the structure is simple, the cost is low, and the portable processing is easy to realize.
Drawings
FIG. 1 is a schematic diagram of the structure of a micro-enzyme thermal biosensor according to the present utility model.
FIG. 2 is a top view of a channel layer of a micro-enzyme thermal biosensor of the present utility model.
FIG. 3 is a schematic view showing the structure of the raised portion of the bottom surface of the channel layer of the micro-enzyme thermal biosensor of the present utility model.
FIG. 4 is a schematic diagram showing the structure of a groove portion of a bottom surface of a channel layer of a micro-enzyme thermal biosensor according to the present utility model.
FIG. 5 is a schematic diagram of the structure of a cover layer of the micro-enzyme thermal biosensor of the present utility model.
FIG. 6 is a schematic view of a cover layer horn inlet of a micro enzyme thermal biosensor of the present utility model.
Fig. 7 is a schematic diagram of the sealing plug structure of the micro enzyme thermal biosensor of the present utility model.
Reference numerals:
1: a channel layer; 2: a cover layer; 3: a sealing plug; 11: recess for engagement with lid, 12: conductive line, 13: liquid flow-through detection channel, 14: signal output electrode, 15: cold side of thermoelectric element (p-side), 16: thermoelectric element hot side (n-side), 17: enzyme modification reaction layer, 18 sample outlet, 19: bottom bump, 110: the side wall extends inwards to form a groove; 21: liquid inlet channel, 22: buffer inlet channel, 23: projections for engagement with the channel layer, 24: semi-cylindrical liquid channel, 25: negative electrode lead outlet, 26: semi-cylindrical buffer liquid channel, 27: positive electrode lead outlet, 28: a horn inlet; 31: sealing ring, 32: annular seal layer, 33: a handle layer.
Detailed Description
The following detailed description of the present utility model is given by way of specific examples, which are given for illustrative purposes only and are not to be construed as limiting the scope of the present utility model. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
The utility model provides a micro enzyme thermal biosensor based on thermoelectric self-energy, which is shown in figure 1 and comprises a channel layer (1), a cover layer (2) arranged above the channel layer and a sealing plug (3) arranged below the channel layer.
The center of the channel layer is a groove-shaped liquid flow detection channel (13), the depth of the channel layer is preferably 3mm, the length of the channel layer is preferably 30mm, and the width of the channel layer is preferably 2mm.
Specifically, the two side walls of the liquid flow detection channel are enzyme modification reaction layers (17), the top wall of the liquid flow detection channel is covered by a cover layer (2), the liquid flow detection channel is connected with the cover layer (2) through a groove (11) embedded with the cover, the sample outlet (18) is connected and sealed with the sealing plug (3), and the liquid flow detection channel is communicated with the outside only through a liquid inlet channel (21) and a buffer liquid inlet channel (22) after the assembly is completed.
Specifically, the thermoelectric elements are all located on the upper surface of the channel layer and form an angle of 90 degrees with the enzyme modification reaction layer, 12-20 pairs are connected in series, the cold end (p end) (15) of each thermoelectric element is connected with the hot end (n end) (16) of each thermoelectric element through a signal output electrode (14), the signal output electrodes (14) are connected through conductive lines (12), and the signal output electrodes are detected through an external voltmeter connected with the conductive lines (12).
Specifically, in order to facilitate magnetron sputtering of the side wall, the junction of the bottom of the liquid flow detection channel of the channel layer and the side wall is detachable and connected with the inward extending groove (110) of the side wall through the bottom bulge (19).
The cover layer comprises a liquid inlet channel (21), a buffer liquid inlet channel (22), a bulge (23) embedded with the channel layer, a negative electrode lead outlet (25) and a positive electrode lead outlet (27).
Specifically, the liquid inlet channel (21) and the buffer liquid inlet channel (22) are horn-shaped inlets (28) which are tightly connected with the semicylindrical liquid channels (24) and the semicylindrical buffer liquid channels (26) below into a whole, wherein the diameter of the upper part of the horn-shaped inlets is preferably 4mm, the diameter of the opening of the lower part is preferably 1mm, the height of the semicylindrical liquid channels is preferably 2mm, and the horn-shaped inlets and the semicylindrical liquid channels are positioned in the center of the covering layer and can be completely embedded into the liquid flow detection channels below.
The sealing plug (3) comprises a sealing ring (31), an annular sealing layer (32) and a handle layer (33). The sealing ring (31) is tightly attached to the annular sealing layer (32) and the sample outlet (18) of the channel layer, so that liquid can be prevented from flowing out or gas can be prevented from entering.
Specifically, in the detection process, liquid to be detected possibly containing a substrate is injected into a liquid inlet channel (21) of the cover layer, enters through a horn-shaped inlet (28), enters a liquid circulation detection channel (13) of the channel layer through a semi-cylindrical liquid channel (24) at the lower part, reacts with enzyme modification reaction layers (17) at the two sides to generate heat, and a thermal signal passes through a thermoelectric element on the upper surface of the channel layer and comprises a cold end (p end) (15) and a hot end (n end) (16) of the thermoelectric element, converts the thermal signal into electromotive force, and is connected with an external voltmeter of a conductive circuit (12) through a signal output electrode (14) to detect. After the detection is finished, the sealing plug (3) is pulled out, buffer solution is injected from the buffer solution inlet channel (22), the liquid flows out from the sample outlet (18) and is washed, and the liquid can be reused after drying.
The micro reagent detection kit is combined with the thermoelectric self-power supply, and the gold film is adopted to fix the enzyme, so that the enzyme thermal biosensor provided by the utility model has the advantages of environmental protection, recycling, small sample consumption, easiness in miniaturized processing and the like.
Claims (5)
1. A miniature enzyme thermal biosensor based on thermoelectric self-energy supply is characterized in that:
the center of the channel layer is a groove-shaped liquid flow detection channel, two side walls of the liquid flow detection channel are enzyme modification reaction layers, wherein the connection part of the bottom of the liquid flow detection channel and the side walls is detachable and is connected with the groove extending inwards through the bulge at the bottom; the sample outlet is connected and sealed with the sealing plug through the groove embedded with the cover and the cover layer; the upper surface is 12-20 pairs of thermoelectric elements connected in series, the cold end of the thermoelectric elements, namely the p end, is connected with the hot end of the thermoelectric elements, namely the n end, through signal output electrodes, the signal output electrodes are connected through conductive lines, and the signal output electrodes are externally connected with a voltmeter through the conductive lines for detection;
the covering layer is arranged above the channel layer and comprises a liquid inlet channel, a buffer liquid inlet channel, a bulge embedded with the channel layer, a negative electrode lead outlet and a positive electrode lead outlet;
the sealing plug is arranged below the channel layer and comprises a sealing ring, an annular sealing layer and a handle layer.
2. The micro-enzyme thermal biosensor according to claim 1, wherein: the cold end, namely the p end, of the thermoelectric element covered on the upper surface of the channel layer is polyethylene dioxythiophene, namely PEDOT, and the hot end, namely the n end, of the thermoelectric element is an Ag2Se film.
3. The micro-enzyme thermal biosensor according to claim 1, wherein: the channel layer signal output electrode is a mask ion sputtering gold electrode; the conductive circuit is made of two identical metal thin wires, and the material of the metal thin wires is platinum wires or copper wires or aluminum wires.
4. The micro-enzyme thermal biosensor according to claim 1, wherein: the liquid inlet channel and the buffer liquid inlet channel of the cover layer are horn-shaped inlets and are tightly connected with the semicylindrical liquid channel and the semicylindrical buffer liquid channel below into a whole, wherein the semicylindrical liquid channel is positioned in the center of the cover layer and can be completely embedded into the liquid circulation detection channel below.
5. The micro-enzyme thermal biosensor according to claim 1, wherein: the sealing plug adopts a rubber plug, and the sealing plug is tightly attached to the sample outlet of the channel layer through the sealing ring and the annular sealing layer, so that liquid outflow or gas inflow can be prevented, and the sealing plug adopts a rubber plug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320842658.1U CN219653039U (en) | 2023-04-17 | 2023-04-17 | Miniature enzyme thermal biosensor based on thermoelectric self-energy supply |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320842658.1U CN219653039U (en) | 2023-04-17 | 2023-04-17 | Miniature enzyme thermal biosensor based on thermoelectric self-energy supply |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219653039U true CN219653039U (en) | 2023-09-08 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202320842658.1U Active CN219653039U (en) | 2023-04-17 | 2023-04-17 | Miniature enzyme thermal biosensor based on thermoelectric self-energy supply |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN219653039U (en) |
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2023
- 2023-04-17 CN CN202320842658.1U patent/CN219653039U/en active Active
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