Efficient adsorption microwell plate based on upright graphene and detection device

Abstract

The utility model relates to the technical field of biochemical analysis, in particular to an efficient adsorption microwell plate based on upright graphene and a detection device. The utility model discloses an efficient adsorption microwell plate based on upright graphene. The efficient adsorption microwell plate comprises a substrate, an upright graphene film attached to the substrate and an interdigital electrode arranged on the upright graphene film. According to the microwell plate, the upright graphene film attached to the substrate serves as a high-flux carrier container and is used for containing a reaction solution and carrying out various biochemical molecule modification reactions, biochemical reactions and photochemical signal detection reactions, and high-speed immunoadsorption can be achieved under the action of the interdigital electrode (driven by an alternating electric field generated); and the reaction incubation time of immunoadsorption is saved.

Description

Technical Field

[0001] The utility model relates to the technical field of biochemical analysis, in particular to a high-efficiency adsorption microporous plate and a detection device based on upright graphene. Background Art

[0002] Vertical graphene refers to graphene nanosheets that grow vertically on the surface of a substrate. Common preparation methods include thermal chemical vapor deposition, plasma enhanced chemical vapor deposition, graphene self-assembly, etc. Among them, thermal chemical vapor deposition is one of the most commonly used preparation methods. In this method, vertically grown graphene can be prepared on a metal catalyst by controlling the diffusion of a metal catalyst and the growth conditions of graphene.

[0003] Upright graphene has the advantage of ultra-large specific surface area due to its unique upright petal morphology and fluffy structure. Due to the characteristics of its π-π conjugated system, it also has the advantages of excellent conductivity, good stability, and easy functional modification, making it a popular and practical sensing interface material and widely used in the construction of new high-performance biochemical sensing methods.

[0004] However, currently, most of the biochemical sensors based on upright graphene focus on utilizing its excellent conductivity and are concentrated in electrochemical signal detection modes. Its advantage of ultra-large specific surface area has not been fully utilized in other types of signal detection modes. Utility Model Content

[0005] The utility model provides a high-efficiency adsorption microporous plate based on upright graphene. The microporous plate uses an upright graphene film attached to a substrate as a high-throughput carrier container to contain reaction solutions, and conduct various biochemical molecule modification reactions, biochemical reactions, and photochemical signal detection reactions. Under the action of interdigital electrodes (generating an alternating electric field drive), high-speed immunoadsorption can be achieved, saving the reaction incubation time of immunoadsorption.

[0006] The technical solution adopted by the utility model to solve its technical problems is:

[0007] A high-efficiency adsorption microporous plate based on upright graphene comprises a substrate, an upright graphene film attached to the substrate, and interdigital electrodes arranged on the upright graphene film.

[0008] Therefore, through the above technical solution, under the action of the interdigitated electrodes (generating AC electric field drive), high-speed immunoadsorption can be achieved through the attached\grown upright graphene, saving the reaction incubation time of immunoadsorption.

[0009] Preferably, the microplate carries 96 micro-interdigitated electrodes and their circuits.

[0010] Preferably, the substrate is quartz glass or an alumina ceramic sheet.

[0011] Preferably, the thickness of the substrate is 0.25 mm.

[0012] Preferably, the heads of the interdigital electrodes are sealed.

[0013] Preferably, the diameter of the electrode circle of the interdigitated electrode is φ6.8 mm, the hole row spacing is 8.8 mm, and the column spacing is 8.9 mm.

[0014] Preferably, the upright graphene film has an upright petal morphology and a fluffy structure.

[0015] In another aspect of the present application, a detection device is provided, comprising the microplate described above.

[0016] Preferably, the protein adsorption capacity of the detection device is > 500 ng / cm 2 .

[0017] The beneficial effects of the utility model are:

[0018] 1. The microplate described in the present application uses quartz glass as a high-throughput carrier container to contain reaction solutions, conduct various biochemical molecule modification reactions, biochemical reactions, and photochemical signal detection reactions, and can achieve high-speed immunoadsorption under the interdigital electrodes (generating an alternating electric field drive), thereby saving the reaction incubation time of immunoadsorption.

[0019] 2. The microplate described in the present application is used for high-throughput accommodation of reaction solutions and perfectly cooperates with commercialized ELISA instruments for high-throughput absorbance measurements. It can accelerate electrode reactions, achieve rapid immune responses, and realize efficient optical detection of enzyme-linked immunosorbent assays. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The utility model is further described below in conjunction with the accompanying drawings and embodiments.

[0021] Figure 1 It is a schematic structural diagram of a high-efficiency adsorption microporous plate based on upright graphene described in the present application;

[0022] Figure 2 It is a schematic diagram of the structure of the detection device described in this application. DETAILED DESCRIPTION

[0023] The following will clearly and completely describe the concept, specific structure and technical effects of the utility model in combination with the embodiments and drawings, so as to fully understand the purpose, characteristics and effects of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of them. Based on the embodiments of the utility model, other embodiments obtained by technicians in this field without creative work are all within the scope of protection of the utility model. In addition, all the connection / connection relationships involved in the patent do not refer to the direct connection of components, but refer to the formation of a better connection structure by adding or reducing connection accessories according to the specific implementation situation. The various technical features in the creation of the utility model can be combined interchangeably without conflicting with each other.

[0024] Reference Figure 1 A high-efficiency adsorption microporous plate based on upright graphene includes a substrate 1, an upright graphene film 2 attached to the substrate 1, and an interdigital electrode 3 arranged on the upright graphene film 2.

[0025] Specifically, the PECVD method is used to grow upright graphene thin film material on the surface of 0.25 mm thick quartz glass. After this process is completed, upright graphene with unique upright petal morphology and fluffy structure is generated (attached) on the upper surface of the quartz glass.

[0026] Compared with conventional two-dimensional materials, upright graphene has a larger specific surface area and can carry more antigens, antibodies, enzymes, nucleic acids, etc.; compared with other conductive materials, upright graphene has the advantage of extremely small surface impedance; compared with metal materials, upright graphene has better bioaffinity and can provide a non-toxic environment. Therefore, the above three advantages of upright graphene can be used to modify its surface with biochemical molecules (antigens, antibodies, nucleic acids, etc.). The graphene (carbon surface) in the microplate chip can be modified and coupled with antibodies and can be used for immunoadsorption of target antigens.

[0027] Therefore, through the above technical solution, under the action of the interdigitated electrodes (AC electric field driven), high-speed immunoadsorption can be achieved through the attached\grown upright graphene, saving the reaction incubation time of immunoadsorption.

[0028] In one embodiment, the microplate carries 96 micro-interdigitated electrodes 3 and their circuits.

[0029] Specifically, the vertical graphene pure material on the whole quartz glass substrate is processed by laser laser engraving or photolithography, and the IDE type electrode (interdigitated electrode) patterning is used as the goal. The vertical graphene (VG) microplate with a complete circuit is initially processed, and the overall size of the microplate is cut out. The size of the quartz glass is 110.1*74.1mm. At this time, a microplate carries 96 micro-interdigitated electrodes and their circuits. The above microplate is set on a 96-hole plate to obtain a 96-type (hole) microporous device, which can be used to place samples to be tested after being embedded in advance.

[0030] In one embodiment, the substrate 1 is made of quartz glass. Specifically, quartz glass is used as the substrate according to actual requirements and application scenarios of whether light transmission is required.

[0031] In one embodiment, the thickness of the substrate 1 is 0.25 mm. The specific thickness of the substrate can be set according to actual needs.

[0032] In one embodiment, the head of the interdigital electrode 3 is sealed. Specifically, the reaction area (the head of the interdigital electrode) is sealed by glue dispensing or injection processing to prevent water leakage when the surface is modified with biochemical molecules, with each micro interdigital electrode head being leak-free.

[0033] Therefore, through the above technical solution, the interdigital electrode will not leak liquid.

[0034] In one embodiment, the diameter of the electrode circle of the interdigital electrode 3 is φ6.8 mm, the hole row spacing is 8.8 mm, and the column spacing is 8.9 mm. The specific arrangement of the interdigital electrodes 3 can be designed according to actual needs; the circuit of the interdigital electrodes can also be designed according to actual needs.

[0035] In another aspect of the present application, a detection device is provided, comprising the microplate described above.

[0036] Specifically, the microplate is a transparent plastic plate used to place samples to be tested, and the plate has multiple rows of small holes of uniform size.

[0037] In one embodiment, the detection device has an adsorption capacity of protein > 500 ng / cm 2 .

[0038] In one embodiment, the structure of the detection device refers to Figure 2 .

[0039] Specifically, the microplate reader is actually a phase-changing photoelectric colorimeter or spectrophotometer, and its basic working principle and main structure are basically the same as those of the photoelectric colorimeter. The light waves emitted by the light source lamp are converted into a beam of monochromatic light through a filter or a monochromator, and enter the sample to be tested in the plastic microplate. Part of the monochromatic light is absorbed by the sample, and the other part is irradiated to the photodetector through the sample. The photodetector converts the light signal of different strengths of the sample to be tested into a corresponding electrical signal. After pre-amplification, logarithmic amplification, analog-to-digital conversion and other signal processing, the electrical signal is sent to the microprocessor for data processing and calculation, and finally the display and printer display the results. The microprocessor also controls the movement of the mechanical drive mechanism in the X and Y directions through the control circuit to move the microplate, thereby realizing the automatic sampling detection process. Other microplate readers use manual movement of the microplate for detection, thus eliminating the mechanical drive mechanism and control circuit in the X and Y directions, making the instrument smaller and simpler in structure.

[0040] A microplate is a transparent plastic plate that has been pre-embedded and is used to place samples to be tested. The plate has multiple rows of small holes of uniform size, and the corresponding antigens or antibodies are embedded in the holes. Each small hole on the microplate can hold a few tenths of a milliliter of solution.

[0041] Therefore, through the above-mentioned technical scheme, the detection device described in the present application can be used to accommodate reaction solutions with high throughput and perfectly adapt to commercialized ELISA instruments for high-throughput absorbance measurement, thereby accelerating electrode reactions, achieving rapid immune responses, and realizing efficient enzyme-linked immunosorbent assay absorbance measurement.

[0042] The specific process of using the microplate is as follows: using the upright graphene film as an adsorption carrier, incubating protein molecules (pure HRP enzyme or HRP enzyme-labeled antibodies) of different concentrations (100ng / mL to 50ug / mL) on the surface of the upright graphene film, incubating at room temperature for 1 hour, and then washing with the prepared detergent after the incubation to remove the unadsorbed or weakly adsorbed protein molecules, and finally performing a TMB color development experiment. Benchmarking the highest adsorption capacity 96-well ELISA plate (Cat. No. 9018, High bingding, the maximum adsorption amount of antibody protein is 400 to 500ng / cm 2 ) At the same protein concentration and after 16 h of incubation, the TMB color development of the upright graphene film is much deeper than that of the ELISA plate. Therefore, it can be determined that the adsorption amount of the upright graphene film is much greater than 500 ng / cm 2 .

[0043] The above is a specific description of the preferred implementation of the utility model, but the invention of the utility model is not limited to the described embodiments. Technical personnel familiar with the field can also make various equivalent deformations or substitutions without violating the spirit of the utility model. These equivalent deformations or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A high-efficiency adsorption microporous plate based on upright graphene, It is characterized in that It comprises a substrate (1), a vertical graphene film (2) attached to the substrate (1), and interdigital electrodes (3) arranged on the vertical graphene film (2).

2. The microplate according to claim 1, It is characterized in that The microporous plate carries 96 micro-interdigitated electrodes (3) and their circuits.

3. The microplate according to claim 1, It is characterized in that The substrate (1) is quartz glass or an alumina ceramic sheet.

4. The microplate according to claim 1, It is characterized in that The thickness of the substrate (1) is 0.25 mm.

5. The microplate according to claim 1, It is characterized in that The heads of the interdigital electrodes (3) are sealed.

6. The microplate according to claim 1, It is characterized in that The electrode circle diameter of the interdigital electrode (3) is φ6.8 mm, the hole row spacing is 8.8 mm, and the column spacing is 8.9 mm.

7. The microplate according to claim 1, It is characterized in that The upright graphene film (2) has an upright petal morphology and a fluffy structure.

8. A detection device, It is characterized in that A microplate comprising any one of claims 1 to 7.

9. The detection device according to claim 8, It is characterized in that The detection device comprises an ELISA instrument.

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