CN219348637U - Magnetic fluorescent microfluidic quantitative detection device - Google Patents
Magnetic fluorescent microfluidic quantitative detection device Download PDFInfo
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- CN219348637U CN219348637U CN202223383106.3U CN202223383106U CN219348637U CN 219348637 U CN219348637 U CN 219348637U CN 202223383106 U CN202223383106 U CN 202223383106U CN 219348637 U CN219348637 U CN 219348637U
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56938—Staphylococcus
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The utility model belongs to the field of detection devices, and particularly relates to a magnetic fluorescent microfluidic quantitative detection device; comprises a cover plate and a base plate; a sample inlet, a sample outlet and a first reaction tank are arranged on the surface of the cover plate; one end of the first reaction tank is communicated with the first reaction channel, and the communicating end is recorded as the upper end of the first reaction tank; the other end of the first reaction channel is communicated with a first sample inlet; the left end of the first reaction tank is communicated with the second sample inlet, the lower end of the first reaction tank is communicated with the third sample inlet, and the right end of the first reaction tank is communicated with a magnet adsorption sheet taking and placing port and a visual detection port; meanwhile, the bottom height of the visual detection port is higher than that of the first reaction tank; a magnet sheet is arranged in the visual detection port, the upper end of the visual detection port is connected with the optical fiber channel, and the right end of the visual detection port is communicated with the sample outlet; the base plate is used for jogging and sealing the butt joint cover plate to form an integrated structure, and the optical fiber channel is arranged between the base plate and the butt joint cover plate. The utility model has the advantages of simple structure, high accuracy, good stability, less sample consumption and short time consumption.
Description
Technical Field
The utility model belongs to the field of detection devices, and particularly relates to a magnetic fluorescent microfluidic quantitative detection device.
Background
Food-borne pathogenic bacteria have the characteristics of wide spread, strong toxicity to human bodies and high pollution frequency, and become the first cause of initiating food safety events. Staphylococcus aureus (abbreviated as staphylococcus aureus) is a common food-borne pathogenic bacterium and widely exists in food raw materials and auxiliary materials and production environments. Based on statistics of authorities, food safety events caused by staphylococcus aureus account for more than 25% of the total safety events of food-borne pathogenic bacteria. In addition, the metabolism of the staphylococcus aureus can produce various human toxins (hemolysin, enterotoxin and the like) and invasive enzymes (plasma coagulase, fibrinolytic enzyme) and the like, so as to cause human pneumonia, pericarditis and even septicemia, wherein the methicillin-resistant staphylococcus aureus has become a third disease infection source after HIV (human immunodeficiency Virus) and HBV (hepatitis B Virus). Therefore, the control of the staphylococcus aureus in the food has important significance.
In recent years, the staphylococcus aureus standard detection method is mainly divided into a plate counting method and a biochemical identification detection method, wherein the plate counting method and the biochemical identification detection method have the defects of long consumption time, large result hysteresis, complex operation, high technical requirements on operators and the like. The latter improves the qualitative speed, but the PCR equipment has the defects of high price, large volume, easy environmental influence, false positive and the like. At present, due to the limitation of complex operation processes such as thallus proliferation, species identification and the like, the quantitative detection time of single-batch staphylococcus aureus usually needs 3-5 days, and the existing staphylococcus aureus detection equipment has high laboratory dependence and strong operation speciality, and cannot meet the requirements of the market on rapid detection of staphylococcus aureus.
Disclosure of Invention
Aiming at the defects of the prior art, the utility model provides the magnetic fluorescence microfluidic quantitative detection device for further improving the accuracy of the detection of the staphylococcus aureus and shortening the detection time, improves the performance of detecting the staphylococcus aureus by a microfluidic technology, and solves the technical problem that the traditional quantitative detection of the staphylococcus aureus is excessively long.
In order to achieve the technical aim, the utility model provides a magnetic fluorescence microfluidic quantitative detection device, which comprises a cover plate, a substrate, a first sample inlet, a second sample inlet, a third sample inlet, a magnet adsorption sheet taking and placing port, a visual detection port, a sample outlet and a fiber channel;
the first sample inlet, the second sample inlet, the third sample inlet, the magnet adsorption sheet taking and placing port, the visual detection port and the sample outlet are all arranged on the surface of the cover plate;
the surface of the cover plate is also provided with a first reaction channel and a first reaction groove; one end of the first reaction groove is communicated with the first reaction channel, and the end, communicated with the first reaction channel, of the first reaction groove is taken as the upper end; the other end of the first reaction channel is connected with a first sample inlet through a channel;
the left end of the first reaction tank is communicated with the sample inlet, and the lower end of the first reaction tank is communicated with the sample inlet in three phases; the right end of the first reaction tank is communicated with a magnet adsorption sheet taking and placing port and a visual detection port; the upper end of the magnet adsorption sheet taking and placing port and the visual detection port are connected with the optical fiber channel, and the right end of the magnet adsorption sheet taking and placing port is communicated with the sample outlet;
the bottom heights of the magnet adsorption sheet taking and placing opening and the visual detection opening are higher than the bottom height of the first reaction tank, the two form a height difference, and the height of a channel between the two is consistent with the bottom heights of the magnet adsorption sheet taking and placing opening and the visual detection opening; so as to prevent the liquid in the first reaction tank from directly flowing to the magnet adsorption sheet taking and placing port and the visual detection port; when the liquid in the first reaction tank is accumulated to a certain height, the liquid can flow to the magnet adsorption sheet taking and placing port and the visual detection port;
the magnet adsorption sheet taking and placing port and the surface of the bottom of the visual detection port are attached with magnet sheets;
the base plate and the cover plate have the same length and width and are used for jogging and sealing the butt joint cover plate to form an integrated structure.
Preferably, the first sample inlet consists of a first sample inlet and a second sample inlet; the second sample inlet consists of a third sample inlet; the sample inlet III consists of a fourth sample inlet and a fifth sample inlet;
the first sample inlet and the second sample inlet are mutually independent and are communicated through a pipeline to form a U-shaped structure, and the bottom end of the U-shaped structure is communicated with the first reaction channel through a channel;
the third sample inlet, the fourth sample inlet and the fifth sample inlet are mutually independent; one end of the first reaction tank is communicated with the first reaction tank through a channel;
more specifically, the third sample inlet is communicated with the left side of the first reaction tank, a pipeline is arranged at the lower end of the first reaction tank, 2 branches are arranged along the extending direction of the pipeline, and the branches are sequentially marked as a first branch and a second branch; the first branch is communicated with the fourth sample inlet, and the second branch is communicated with the fifth sample inlet;
preferably, the bottoms of the magnet adsorption sheet taking and placing opening and the visual detection opening are higher than the bottom of a channel connecting the magnet adsorption sheet taking and placing opening and the first reaction tank, and the height difference is 0.5-0.8 cm.
Preferably, the thickness of the magnet sheet is 0.1-0.2 cm, and the area of the magnet sheet is matched with the area of the detection port (10).
Preferably, the first reaction channel is a serpentine channel.
Preferably, the cover plate is a cuboid of 6cm×5cm×1cm, the substrate is a cuboid of 6cm×5cm×0.5cm, and the materials are polydimethylsiloxane;
preferably, the first sample inlet, the second sample inlet, the third sample inlet, the magnet adsorption sheet taking and placing port, the visual detection port and the sample outlet are all arranged on the surface of the cover plate and penetrate through the cover plate;
the optical fiber channel is arranged between the cover plate and the base plate.
Compared with the prior art, the utility model has the following advantages and effects:
(1) The magnetic fluorescence micro-flow quantitative control detection device provided by the utility model improves the performance of detecting the staphylococcus aureus by a micro-flow control technology, solves the problems of long time consumption, high separation and purification difficulty, complex operation and other pain points in the traditional quantitative detection of the staphylococcus aureus, and has the advantages of short time consumption, high accuracy, good stability, small sample consumption volume and the like.
(2) The utility model has simple structure and operation flow, and is suitable for industrial production.
(3) The device based on the utility model realizes rapid detection, only needs about 30 minutes from the construction of the sensor to the completion of sample analysis, breaks through the problem that the traditional method for detecting the staphylococcus aureus needs several days, can realize rapid and quantitative detection, and has extremely important significance in the fields of food safety detection and the like.
(4) The utility model basically realizes automation of operation, and solves the problem of complex operation of the traditional method.
Drawings
FIG. 1 is a schematic diagram of the overall structure of a magnetic fluorescence microfluidic quantitative detection device;
FIG. 2 is a schematic diagram of a cover plate structure;
wherein: 1-a cover plate; 2-a substrate; 3-a first sample inlet; 4-a second sample inlet; 5-a third sample inlet; 6-a fourth sample inlet; 7-a fifth sample inlet; 8-a first reaction channel; 9-a first reaction tank; 10-a magnet adsorption sheet taking and placing port and a visual detection port; 11-fibre channel; 12-sample outlet.
Detailed description of the preferred embodiments
In order that the above-recited objects, features and advantages of the present utility model will become more readily apparent, a more particular description of the utility model will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
Example 1
As shown in fig. 1-2, the device comprises a cover plate 1, a substrate 2, a first sample inlet 3, a second sample inlet 4, a third sample inlet 5, a fourth sample inlet 6, a fifth sample inlet 7, a first reaction channel 8, a first reaction tank 9, a magnet adsorption sheet taking and placing port, a visual detection port 10, an optical fiber channel 11 and a sample outlet 12;
the first sample inlet 3, the second sample inlet 4, the third sample inlet 5, the fourth sample inlet 6, the fifth sample inlet 7, the magnet adsorption sheet taking and placing port, the visual detection port 10 and the sample outlet 12 are all arranged on the surface of the cover plate 1 and all penetrate through the cover plate 1;
the surface of the cover plate 1 is also provided with a first reaction channel 8 and a first reaction groove 9; one end of the first reaction groove 9 is communicated with the first reaction channel 8, and the end of the first reaction groove 9 communicated with the first reaction channel 8 is taken as the upper end; the other end of the first reaction channel 8 is connected with a first sample inlet through a channel; the first reaction channel 8 is a serpentine channel, so that the solution is fully mixed in the channel;
the third sample inlet 5 is communicated with the left side of the first reaction tank 9, a pipeline is arranged at the lower end of the first reaction tank, and 2 branches are arranged along the extending direction of the pipeline and are sequentially marked as a first branch and a second branch; the first branch is communicated with the fourth sample inlet 6, and the second branch is communicated with the fifth sample inlet 7; the right end of the first reaction tank 9 is communicated with a magnet adsorption sheet taking and placing port and a visual detection port 10; the upper end of the magnet adsorption sheet taking and placing port and the visual detection port 10 is connected with a fiber channel 11, and the right end is communicated with a sample outlet 12;
the surface of the bottom of the magnet adsorption sheet taking and placing opening and the surface of the bottom of the visual detection opening 10 are provided with magnet sheets with the thickness of 0.1cm, and the area of the magnet sheets is matched with the area of the bottom of the magnet adsorption sheet taking and placing opening and the surface of the bottom of the visual detection opening 10; the bottom of the magnet adsorption sheet taking and placing opening and the visual detection opening 10 is higher than the bottom of a channel connecting the magnet adsorption sheet taking and placing opening and the visual detection opening with the first reaction tank 9, the height difference of the magnet adsorption sheet taking and placing opening and the visual detection opening is 0.5cm, and the height of the channel between the magnet adsorption sheet taking and placing opening and the visual detection opening is consistent with the height of the bottom of the magnet adsorption sheet taking and placing opening and the visual detection opening 10; the purpose is to make the sample to be tested have enough reaction time when entering the first reaction tank 9, when the liquid level of the solution in the first reaction tank 9 reaches the height of the channel, the sample can flow into the bottom of the magnet adsorption sheet taking and placing port and the visual detection port 10 along the channel, and at the moment, the sensor after the magnet adsorption reaction attached to the bottom of the magnet adsorption sheet taking and placing port and the visual detection port 10 is utilized to achieve the effect of separating and purifying the staphylococcus aureus;
the cover plate 1 is a cuboid with the thickness of 6cm multiplied by 5cm multiplied by 1cm, the substrate 2 is a cuboid with the thickness of 6cm multiplied by 5cm multiplied by 0.5cm, and the materials are polydimethylsiloxane; the length and width dimensions of the base plate 2 and the cover plate 1 are consistent, and the base plate and the cover plate 1 are used for jogging and sealing the butt joint cover plate 1 to form an integrated structure;
when the base plate 2 is used for embedding and sealing the butt joint cover plate 1, the sealing is equivalent to sealing the openings at one side of the first sample inlet 3, the second sample inlet 4, the third sample inlet 5, the fourth sample inlet 6, the fifth sample inlet 7, the magnet adsorption sheet taking and placing opening, the visual detection opening 10 and the sample outlet 12; the fibre channel 11 is provided between the cover plate 1 and the base plate 2.
The method for detecting the staphylococcus aureus based on the device comprises the following specific operation steps:
s1, synthesizing magnetic nano Material (MN)
Weigh 0.02mol FeCl 3 ·6H 2 O and 0.01mol FeCl 2 ·4H 2 O is respectively dissolved in 100mL of ultrapure water, deoxidization is carried out under the nitrogen atmosphere at 80 ℃, 25% ammonia water is added dropwise, the pH value of the solution is adjusted to 10.0, meanwhile, the solution is continuously stirred in the whole reaction process, the reaction is stopped after 4 hours, the reaction solution is naturally cooled to room temperature, then the reaction solution is kept stand for 4 hours, and the filtered precipitate is dried for 24 hours at 70 ℃ after being washed with ethanol and ultrapure water for three times, so as to obtain the magnetic nano material.
S2, synthetic fluorescent material (AuNCs)
20mL of 4mM HAuCl was removed 4 Mixing with 20mL of 6mM reduced glutathione, stirring and heating at 90 ℃ for 6 hours, cooling the obtained reaction liquid to room temperature, and obtaining the fluorescent material, wherein the fluorescent material is stored at 4 ℃ in a dark place for further use.
S3, preparing MN@PEI composite material
Transferring 10mL of 1g/mL of MN solution, adding 40mL of polyethyleneimine solution with mass concentration of 0.5%, incubating for 2 hours, centrifuging, taking precipitate, and centrifugally washing with ultrapure water for 3 times, wherein the washed precipitate is the MN@PEI composite material;
re-dissolving the obtained MN@PEI composite material into ultrapure water to obtain an MN@PEI composite material aqueous solution with the mass concentration of 1 g/mL;
s4, enabling 0.3mL of MN@PEI composite material aqueous solution to flow to a first reaction channel 8 at 30 mu L/min through a first sample inlet 3 by using a syringe pump, simultaneously enabling 0.3mL of fluorescent material (AuNCs) to flow to the first reaction channel 8 at 30 mu L/min through a second sample inlet 4 by using a syringe pump, controlling the magnetic material and the fluorescent material to be mixed in the first reaction channel 8, enabling the mixture to flow to a first reaction tank 9 through the channel, and carrying out mixed reaction on the two solutions to obtain the MN@PEI@AuNCs composite material;
s5, enabling 0.3mL of staphylococcus aureus antibody (Ab) water solution with the mass concentration of 10 mug/mL to flow to a first reaction tank 9 through a fourth sample inlet 6 by using a syringe pump at the speed of 200 mug/min, simultaneously enabling a fluorescence extinguishing agent to flow to the first reaction tank 9 through a fifth sample inlet 7 by using the syringe pump at the speed of 200 mug/min, controlling the mixture of the antibody and the fluorescence extinguishing agent and the MN@PEI@AuNCs composite material in the first reaction tank 9, and incubating for 10min to obtain an antibody-magnetic fluorescence composite material solution, namely the quantitative detection sensor based on a magnetic fluorescence nano system.
S6 0.3mL of c=10 was added dropwise with a syringe pump at 200 μl/min through the third inlet 5 -8 The method comprises the steps of controlling a bacterial liquid to flow to a first reaction tank 9 through a channel and be mixed with a quantitative detection sensor to obtain a sample-sensor system to be detected, incubating for 10min, continuously introducing ultrapure water to enable the liquid level of a system to reach the heights of the bottom surfaces of a magnet adsorption sheet taking and placing port and a visual detection port 10, then enabling liquid in the first reaction tank 9 to flow to the magnet adsorption sheet taking and placing port and the visual detection port 10, obtaining a purified sample-sensor system to be detected by utilizing the magnet adsorption sheet taking and placing port and the magnet sheet at the bottom surface of the visual detection port 10, controlling the system to flow to the magnet adsorption sheet taking and placing port and the visual detection port 10 through the channel, and measuring a fluorescence signal F= 297.2948 of the system by laser irradiating from an optical fiber channel 11 under a three-purpose ultraviolet lamp from the position of the magnet adsorption sheet taking and placing port and the visual detection port 10; the standard curve is constructed through the fluorescence signal and the corresponding concentration, so that the quantitative detection of staphylococcus aureus is realized.
Description: the above embodiments are only for illustrating the present utility model and not for limiting the technical solution described in the present utility model; thus, while the utility model has been described in detail with reference to the various embodiments described above, it will be understood by those skilled in the art that the utility model may be modified or equivalents; all technical solutions and modifications thereof that do not depart from the spirit and scope of the present utility model are intended to be included in the scope of the appended claims.
Claims (9)
1. The magnetic fluorescent microfluidic quantitative detection device is characterized by comprising a cover plate (1), a substrate (2), a first sample inlet, a second sample inlet, a third sample inlet, a magnet adsorption sheet taking and placing port, a visual detection port (10), a sample outlet (12) and an optical fiber channel (11);
the first sample inlet, the second sample inlet, the third sample inlet, the magnet adsorption sheet taking and placing port, the visual detection port (10) and the sample outlet (12) are all arranged on the surface of the cover plate (1);
the surface of the cover plate (1) is also provided with a first reaction channel (8) and a first reaction groove (9); one end of the first reaction groove (9) is communicated with the first reaction channel (8), and the end of the first reaction groove (9) communicated with the first reaction channel (8) is taken as the upper end; the other end of the first reaction channel (8) is connected with a first sample inlet through a channel;
the left end of the first reaction tank (9) is communicated with the sample inlet, and the lower end of the first reaction tank (9) is communicated with the sample inlet in three phases; the right end of the first reaction tank (9) is communicated with a magnet adsorption sheet taking and placing port and a visual detection port (10); the upper end of the magnet adsorption sheet taking and placing port and the visual detection port (10) is connected with the optical fiber channel (11), and the right end is communicated with the sample outlet (12);
the bottom heights of the magnet adsorption sheet taking and placing opening and the visual detection opening (10) are higher than the bottom height of the first reaction tank (9), the two form a height difference, and the height of a channel between the two is consistent with the bottom heights of the magnet adsorption sheet taking and placing opening and the visual detection opening (10); so as to prevent the liquid in the first reaction tank (9) from directly flowing to the magnet adsorption sheet taking and placing port and the visual detection port (10); when the liquid in the first reaction tank (9) is accumulated to a certain height, the liquid can flow to the magnet adsorption sheet taking and placing port and the visual detection port (10);
the magnet adsorption sheet taking and placing port and the visual detection port (10) are provided with a magnet sheet on the bottom surface;
the base plate (2) and the cover plate (1) are consistent in length and width and used for being embedded and sealed with the cover plate (1) to form an integrated structure.
2. The magnetic fluorescence microfluidic quantitative detection device according to claim 1, wherein the first sample inlet consists of a first sample inlet (3) and a second sample inlet (4); the second sample inlet consists of a third sample inlet (5); the third sample inlet consists of a fourth sample inlet (6) and a fifth sample inlet (7);
the first sample inlet (3) and the second sample inlet (4) are mutually independent and are communicated through a pipeline to form a U-shaped structure, and the bottom end of the U-shaped structure is communicated with the first reaction channel (8) through a channel;
the third sample inlet (5), the fourth sample inlet (6) and the fifth sample inlet (7) are arranged independently; one end of the reaction tank is communicated with the first reaction tank (9) through a channel.
3. The magnetic fluorescence microfluidic quantitative detection device according to claim 2, wherein the third sample inlet (5) is communicated with the left side of the first reaction tank (9), a pipeline is arranged at the lower end of the first reaction tank (9), and 2 branches are arranged along the extending direction of the pipeline and are sequentially marked as a first branch and a second branch; the first branch is communicated with a fourth sample inlet (6), and the second branch is communicated with a fifth sample inlet (7).
4. The magnetic fluorescent microfluidic quantitative detection device according to claim 1, wherein the bottoms of the magnet adsorption sheet taking and placing port and the visual detection port (10) are higher than the bottom of a channel connecting the magnet adsorption sheet taking and placing port and the first reaction tank (9), and the height difference is 0.5-0.8 cm.
5. The magnetic fluorescent microfluidic quantitative detection device according to claim 1, wherein the thickness of the magnet sheet is 0.1-0.2 cm, and the area of the magnet sheet is matched with the area of the detection port (10).
6. The magnetic fluorescent microfluidic quantitative detection device according to claim 1, wherein the first reaction channel (8) is a serpentine channel.
7. The magnetic fluorescent microfluidic quantitative detection device according to claim 1, wherein the cover plate (1) is a cuboid of 6cm×5cm×1cm, the substrate (2) is a cuboid of 6cm×5cm×0.5cm, and the materials are polydimethylsiloxane.
8. The magnetic fluorescent microfluidic quantitative detection device according to claim 1, wherein the first sample inlet, the second sample inlet, the third sample inlet, the magnet adsorption sheet taking and placing port, the visual detection port (10) and the sample outlet (12) are all arranged on the surface of the cover plate (1), and all penetrate through the cover plate (1).
9. The magnetic fluorescent microfluidic quantitative detection device according to claim 1, wherein the optical fiber channel (11) is arranged between the cover plate (1) and the base plate (2).
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202223383106.3U CN219348637U (en) | 2022-12-16 | 2022-12-16 | Magnetic fluorescent microfluidic quantitative detection device |
| DE202023101513.1U DE202023101513U1 (en) | 2022-12-16 | 2023-03-27 | Magnetic, fluorescent, microfluidic and quantitative detection device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202223383106.3U CN219348637U (en) | 2022-12-16 | 2022-12-16 | Magnetic fluorescent microfluidic quantitative detection device |
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| Publication Number | Publication Date |
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| CN219348637U true CN219348637U (en) | 2023-07-14 |
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| CN202223383106.3U Active CN219348637U (en) | 2022-12-16 | 2022-12-16 | Magnetic fluorescent microfluidic quantitative detection device |
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|---|---|
| CN (1) | CN219348637U (en) |
| DE (1) | DE202023101513U1 (en) |
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| DE202023101513U1 (en) | 2023-06-07 |
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