Terahertz detection biomolecule chip for solution environment and manufacturing method thereof

Abstract

The present application relates to a kind of terahertz detection biomolecule chip for solution environment and manufacturing method, by being equipped with the nanometer hole of aperture uniformity in inner cavity, after being powered on to biomolecule chip, the biological macromolecule existing in biological solution can move from other positions in inner cavity to the end where nanometer hole is located.Different biological macromolecule selects different aperture nanometer hole nanometer hole, and single biological macromolecule in conformity with the size of the size of the single biological macromolecule opened by itself is screened, so as to limit and locate single biological macromolecule in nanometer hole, avoid biological macromolecule to experience fast and various conformational transition in biological solution.Compared with prior art, the present application discloses a kind of terahertz detection biomolecule chip for solution environment and manufacturing method, can realize the purpose of in-situ real-time imaging detection of single biological macromolecule in solution state.

Description

Technical Field

[0001] This invention relates to the field of micro-nano sensing and detection technology, and in particular to a terahertz detection biomolecular chip for solution environments and its manufacturing method. Background Technology

[0002] Terahertz (THz) waves lie between infrared and microwaves, possessing unique properties not found in other wavebands. The vibrational and rotational energy levels of fundamental biological macromolecules such as proteins and nucleic acids fall within the terahertz band, making terahertz waves a natural and effective tool for detecting the motion of biological macromolecules. However, due to the relatively long wavelength of terahertz waves (between 30 micrometers and 3 millimeters), the detection of the conformation and dynamics of biological macromolecules using terahertz waves is limited by spatial resolution. Terahertz near-field imaging technology can overcome the spatial resolution limitations of terahertz wave imaging, and the imaging resolution for semiconductor devices and dry biological organisms has now reached the tens of nanometers level.

[0003] Unfortunately, current terahertz near-field imaging research mainly focuses on dry-state, biomolecular / cell population levels, lacking corresponding methods for detecting active single molecules / single cells under physiological conditions, which is urgently needed. Under normal physiological conditions, water not only strongly absorbs terahertz waves but also contaminates terahertz near-field nanoprobes. Furthermore, biomolecules in solution undergo rapid and diverse conformational changes, posing challenges for in-situ real-time imaging. Therefore, single-molecule terahertz imaging detection of biomolecules in solution remains an unsolved problem both domestically and internationally. Summary of the Invention

[0004] To address the limitations of in-situ real-time terahertz imaging detection of individual biomolecules in solution environments due to the strong absorption of terahertz waves by water and the rapid and diverse conformational changes that biomolecules undergo, this invention proposes a terahertz biomolecule detection chip for solution environments.

[0005] The technical solution adopted in this invention is a terahertz detection biomolecule chip for solution environment, comprising a chip body, an inner cavity communicating with the outside of the chip body, a uniformly sized nanopore penetrating the inner cavity and capable of accommodating a single biomolecule, and a detection window made of a conductive material that can be penetrated by terahertz waves attached to the outside of the nanopore.

[0006] Preferably, the pore size of the nanopores ranges from 10 nanometers to 50 nanometers.

[0007] Preferably, the depth of the nanopores ranges from 10 nanometers to 50 nanometers.

[0008] Preferably, the nanopores are circular, and multiple nanopores are distributed in an array.

[0009] Preferably, the inner cavity is connected to the outside through a microfluidic channel, the pore size of which ranges from 10 micrometers to 1000 micrometers.

[0010] Preferably, the chip body includes a support layer, a substrate, and a bottom cover that are sequentially bonded together. The substrate and the support layer form an inner cavity. The support layer has nanopores, and the bottom cover is provided with microfluidic channels.

[0011] Preferably, the height of the inner cavity is between 150 micrometers and 600 micrometers.

[0012] Preferably, the chip body is connected to or etched with an oscillation circuit for controlling the power supply switching and / or an intelligent control circuit for controlling the current flow.

[0013] To address the limitations of in-situ real-time terahertz imaging detection of individual biomolecules in solution environments due to the strong absorption of terahertz waves by water and the rapid and diverse conformational changes that biomolecules undergo, this invention also proposes a method for manufacturing a biomolecular chip.

[0014] The technical solution adopted in this invention is a method for manufacturing a biomolecular chip, comprising:

[0015] Etch the internal cavity onto the chip body;

[0016] Nanopores with uniform pore size and connected to the internal cavity are fabricated on the chip body;

[0017] The detection window is moved to the location of the nanopore.

[0018] Preferred options also include:

[0019] The polished and ultrasonically cleaned substrate is placed in a furnace tube, and a PECVD system is used with silane, ammonia and nitrogen as gas sources. Temperature, airflow and reaction time are controlled to deposit a silicon nitride support layer with a thickness of 20 to 30 nanometers on the substrate surface.

[0020] On the back side of a substrate with a silicon nitride support layer deposited, the inner cavity is etched out using a KOH wet etching process;

[0021] At least two nanopores or nanopore arrays with different pore sizes were fabricated on a silicon nitride support layer using focused helium ion beam technology. The pore size ranged from 10 nanometers to 50 nanometers.

[0022] A PMMA-assisted wet transfer method was used to transfer graphene films with fewer than 5 layers prepared by CVD to the nanopores on a silicon nitride support layer.

[0023] The substrate that has been polished and ultrasonically cleaned is used as the bottom cover. Holes are drilled at two intervals on the bottom cover by mechanical processing, which serve as the liquid inlet and the liquid outlet, respectively.

[0024] The unetched portion of the lower surface of the substrate and the upper surface of the bottom cover are subjected to oxygen plasma surface treatment, and then the two are bonded together at room temperature.

[0025] Metallic gold electrodes are electrochemically deposited on the edge of the graphene detection window.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] This application discloses a terahertz detection biomolecule chip for solution environments. By incorporating uniformly sized nanopores within the chip's internal cavity, biomolecules present in the biological solution can move from other locations within the cavity to the nanopores after the chip is energized. The nanopores screen individual biomolecules that match their size, thereby confining and positioning them within the nanopores and preventing them from undergoing rapid and diverse conformational changes in the biological solution. Furthermore, since the confined and positioned biomolecules are located within the nanopores, and a detection window made of a conductive material permeable to terahertz waves is attached to the nanopore, the detection window acts as an insulating layer during scanning imaging of the biomolecules, protecting the terahertz nanoprobe from contact with the biological solution and preventing the strong absorption of terahertz waves by water from affecting the scanning imaging.

[0028] Compared with existing technologies, the terahertz near-field imaging detection biomolecule chip disclosed in this application can achieve the purpose of terahertz in-situ real-time imaging detection of individual biomolecules in solution.

[0029] This application also discloses a method for manufacturing a biomolecular chip, which involves etching an inner cavity on the chip body; preparing at least two nanopores with uniform pore size and communicating with the inner cavity on the chip body; and transferring the detection window to the location of the nanopore.

[0030] Compared with the prior art, the method for manufacturing a biomolecular chip disclosed in this application can achieve the purpose of terahertz in-situ real-time imaging detection of individual biomacromolecules in solution. Attached Figure Description

[0031] The present invention will now be described in detail with reference to the embodiments and accompanying drawings, wherein:

[0032] Figure 1 This is a schematic diagram of the biomolecular chip structure of the present invention;

[0033] Figure 2This is a schematic diagram of the biomolecular chip fabrication process of the present invention;

[0034] Figure 3 This is a top view schematic diagram of the silicon nitride support layer with nanopores of different pore sizes etched according to the present invention;

[0035] Figure 4 This is a schematic diagram of the biomolecular chip of the present invention used for terahertz near-field imaging detection of single biomolecules.

[0036] Label Explanation:

[0037] 1. Substrate; 2. Groove; 3. Silicon nitride support layer; 4. Nanopore; 5. Graphene window; 6. Cap; 7. Inlet; 8. Outlet; 9. Gold electrode; 10. External power supply; 11. Confined biomolecule; 12. Free biomolecule; 13. Terahertz source; 14. Terahertz detector; 15. Nano probe; 16. Tuning fork. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of the present invention clearer, the embodiments of the present invention will be further described in detail below with reference to the accompanying drawings. Examples of embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar components or components having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0039] This invention discloses a terahertz detection biomolecule chip for use in solution environments. Please refer to [link / reference]. Figures 1 to 4 The chip includes a chip body with an internal cavity that communicates with the outside. The surface of the chip body has at least two nanopores 4 with uniform pore size that can accommodate a single biomolecule, which penetrate into the internal cavity. A detection window made of a conductive material that can be penetrated by terahertz waves is attached to the outside of the nanopores 4.

[0040] By providing at least two uniformly sized nanopores 4 within the inner cavity, biomolecules present in the biological solution can move from other locations within the inner cavity to the end where the nanopores 4 are located after the biomolecule chip is energized. The nanopores 4 screen individual biomolecules that match their opening size, thereby confining and positioning the individual biomolecules within the nanopores 4 and preventing them from undergoing rapid and diverse conformational changes in the biological solution. Furthermore, since the confined and positioned biomolecules are located within the nanopores 4, and a detection window is attached to the nanopores 4, the detection window is made of a conductive material that can be penetrated by terahertz waves. Therefore, during scanning imaging of the biomolecules, the detection window can act as an isolation layer to protect the terahertz nanoprobe 15 from contact with the biological solution, thus preventing the strong absorption of terahertz waves by water from affecting the scanning imaging of the terahertz nanoprobe 15. Compared with the prior art, the terahertz near-field imaging detection biomolecule chip disclosed in this application can achieve the purpose of in-situ real-time terahertz imaging detection of individual biomolecules in solution.

[0041] It should be noted that this application uses the nanopore 4 as a nanochannel, which can not only provide a normal physiological environment for biomacromolecules, but also provide a cell-like confined physical environment for biomacromolecules. The size of the nanopore 4 allows for precise positioning of individual biomacromolecules, and the combination of the local electric field of the nanopore 4 under cross-mode voltage to manipulate biomacromolecules is beneficial for the capture of individual macromolecules and for the terahertz near-field imaging detection and analysis of biomacromolecules.

[0042] Specifically, for the detection of different biomolecules, since biomolecules vary in size, the pore size of the nanopore 4 can be selected differently. Generally, it only needs to be about 5 nm larger than the size of the biomolecule. Due to the confinement effect of the nanopore, it can be ensured that each nanopore contains at most one biomolecule. Using multiple nanopores 4 with uniform pore size can increase the capture probability of biomolecules, thereby obtaining multiple control groups for convenient detection and analysis.

[0043] The detection window needs to be made of a conductive material that can be penetrated by terahertz waves. This is because the terahertz waves emitted by the external terahertz source 13 need to penetrate the detection window to act on the biomolecules and resonate with them. The conductive material is required because the positive or negative terminal of the external power supply needs to be connected to the detection window, allowing the biomolecules to move closer to the nanopore 4 and be captured by it. The device used with the biomolecule chip can include a power supply, with one terminal connected to the detection window and the other connected to the inner cavity.

[0044] In addition, the inner cavity needs to be connected to the outside, and the location where it is connected to the outside is used to add the biological solution. In some other embodiments, the inner cavity can be open, and the open side of the biomolecular chip can be directly immersed in the biological solution when in use.

[0045] In some embodiments, the pore size of the nanopore 4 ranges from 10 nanometers to 50 nanometers.

[0046] Specifically, biological macromolecules include substances such as nucleic acids and proteins. Nucleic acids are relatively small, about a few nanometers in size, while proteins are relatively large, about tens of nanometers in size. Therefore, setting the pore size of nanopore 4 between 10 nanometers and 50 nanometers allows for the detection of both small nucleic acids and large proteins.

[0047] In some more specific embodiments, since most nucleic acids are only a few nanometers in size, multiple nanopores 4 in the nucleic acid detection array are arranged in a normal distribution on the chip body with an average size of 2.5 nanometers. That is, between 1 nanometer and 10 nanometers, the closer the size of the nanopore 4 is to 2.5 nanometers, the more nanopores 4 are arranged. In addition to nanopores 4 with uniform pore size, nanopores 4 with different pore sizes can also be provided, thereby realizing the capture and detection of biomolecules of different sizes.

[0048] In some embodiments, the depth of the nanopore 4 ranges from 10 nanometers to 50 nanometers.

[0049] Specifically, to ensure that biomolecules are also contained within the nanopore 4 along their length, the depth of the nanopore 4 is limited to between 10 and 50 nanometers. This allows for better capture of biomolecules, facilitating their in-situ placement and improving imaging detection results.

[0050] It should be noted that biomolecules can have shapes such as elongated, rod-shaped, and elliptical. When detecting elongated biomolecules, the nanopore 4 can be designed so that the captured elongated shape is perpendicular to the opening direction of the nanopore 4, making it easier to capture elongated biomolecules.

[0051] In some embodiments, the nanopores 4 are circular, and multiple nanopores 4 are arranged in an array.

[0052] It should be noted that the nanopores 4 are circular. This is because circular nanopores 4 can more effectively prevent biomolecules from getting stuck inside them. The smoother circular shape also reduces the likelihood of this getting stuck. Furthermore, the multiple nanopores 4 are arranged in an array. An array distribution, compared to a discrete distribution, is more conducive to improving the capture probability of the nanopores 4 and thus increasing the working efficiency of the biomolecular chip.

[0053] In some embodiments, the inner cavity is connected to the outside via a microfluidic channel, the pore size of which ranges from 10 micrometers to 1000 micrometers.

[0054] It should be noted that the inner cavity is connected to the outside via microfluidic channels. By limiting the size of the microfluidic channels, the flow rate between the biological solution inside the inner cavity and the external biological solution can be effectively reduced. This prevents a large number of biomolecules from accumulating in the external biological solution to the nanopore 4 under the influence of the applied electric field, thus avoiding excessive congestion of the inner cavity and hindering the capture of individual biomolecules. Furthermore, the microfluidic channels also prevent excessive current generated by the power source from escaping to the outside of the inner cavity, thereby reducing the electric field strength and reducing the capture efficiency.

[0055] Specifically, the selected microfluidic channel has an aperture size ranging from 10 micrometers to 1000 micrometers, which is very small relative to the size of the entire biomolecular chip, while still being able to allow a molecular pump or syringe to inject biological solutions into the inner cavity through the microfluidic channel.

[0056] Furthermore, when the pore size of the microfluidic channel is between 10 micrometers and 1000 micrometers, due to the surface tension of the liquid, the biological solution in the inner cavity will not flow out of the microfluidic channel without external interference, thereby achieving the purpose of restricting the outflow of the biological solution, which in turn allows the biomolecular chip to be used in a variety of application scenarios.

[0057] In some specific embodiments, the chip body includes a support layer, a substrate 1 and a bottom cover that are sequentially bonded together. The substrate 1 and the support layer form an inner cavity. The support layer has nanopores 4 and the bottom cover is provided with microfluidic channels.

[0058] It should be noted that the chip body includes a support layer, a substrate 1, and a bottom cover that are attached in sequence. In other embodiments, the three can be made of the same material. However, in this embodiment, by setting them separately, the biomolecular chip can be made easier to manufacture. On the other hand, and more importantly, the operator can choose different materials to manufacture the support layer, substrate 1, and bottom cover according to factors such as the required size of the nanopores 4, the required structural strength, and the chemical properties of the biological solution.

[0059] In some specific embodiments, the height of the cavity is characterized by being between 150 micrometers and 600 micrometers.

[0060] Specifically, to achieve a faster response speed for the biomolecular chip, the height of the cavity is set between 150 and 600 micrometers, meaning the maximum distance a biomolecule can travel is 600 micrometers. This allows the nanopore 4 to capture biomolecules more quickly. Setting the minimum cavity height to 150 micrometers ensures that the cavity stores an appropriate amount of biomolecules, which also contributes to the faster capture speed of the biomolecules by the nanopore 4.

[0061] In some specific embodiments, the chip body is connected to or etched with an oscillation circuit for controlling the power supply switching and / or an intelligent control circuit for controlling the current flow.

[0062] The chip body has an oscillation circuit connected or etched on it to control the power supply. The purpose of the oscillation circuit is to allow the biomolecules to move and oscillate in the biological solution in the inner cavity by switching the circuit on and off when the nanopore 4 does not capture the biomolecules. This allows the nanopore 4 to successfully capture the biomolecules that need to be imaged and detected.

[0063] Intelligent control circuits that control the direction of current flow are connected or etched on the chip body. The reason for setting up intelligent control circuits is that different types of biomolecules carry different positive and negative charges, so it is necessary to change the direction of the current in order to make the biomolecules move one end of the nanopore 4.

[0064] Obviously, etching the oscillation circuit and / or intelligent control circuit onto the chip body will result in a higher degree of integration of the biomolecular chip, while connecting it will reduce its cost.

[0065] In other embodiments, increasing the voltage can accelerate the capture rate of individual biomolecules.

[0066] In some embodiments, the detection window is preferably made of graphene. Single-layer or few-layer graphene has significant advantages as a terahertz near-field imaging window: graphene has atomic-level flatness, high terahertz wave transmittance (single-layer graphene transmittance: 97.7%, 2-10 layer graphene transmittance: >80%), and stable physical adsorption capacity for biomacromolecules. This ensures that the protein single molecules adsorbed on the lower surface of graphene can interact with the radiated terahertz near-field signal. At the same time, it can also provide a smooth scanning plane for the terahertz near-field nanoprobe 15, avoiding contamination of the nanoprobe 15 by biological solutions and the influence on the vibration of the nanoprobe 15.

[0067] In other embodiments, substrate 1 may be made of silicon or other semiconductor materials. The support layer is made of a non-conductive material that does not react with the biological solution and can have higher structural strength relative to the material of substrate 1.

[0068] In a particularly detailed embodiment, such as Figure 1 The biomolecular chip shown includes:

[0069] Base 1, with an inner cavity 2 engraved on base 1;

[0070] The silicon nitride support layer 3 has nanopores 4 of varying sizes arranged in a linear pattern; the silicon nitride support layer 3 is fixedly connected to the substrate 1.

[0071] Furthermore, nanopores 4 have different pore sizes, which can confine biomolecules of different sizes;

[0072] Graphene detection window 5 is transferred to silicon nitride support layer 3 by wet transfer.

[0073] The bottom cover 6 has a microfluidic channel including an inlet 7 and an outlet 8; the bottom cover 6 is fixedly connected to the substrate 1; the bottom cover 6 is made of one of silicon, silicon dioxide and polymer.

[0074] Furthermore, the polymeric material includes one of polydimethylsiloxane, polystyrene, polycarbonate, and polymethyl phenylformate.

[0075] Metallic gold electrode 9, which is plated on the graphene detection window 5;

[0076] An external power source 10 has its negative electrode connected to a gold electrode 9, and its positive electrode inserted into the biological solution through an inlet 7. Due to the different charges of biological macromolecules, the positive and negative electrodes of the external power source may also be connected in the opposite way.

[0077] Confined biomacromolecules 11 and free biomacromolecules 12 are present. The biomacromolecules are introduced into the microfluidic channel in solution form through the inlet 7 via a molecular pump or syringe. Under the action of an external electric field, the biomacromolecules move towards the nanopore 4 and are captured by the nanopore 4, forming a small portion of confined biomacromolecules 11. These are the biomacromolecules that can be detected by terahertz near-field imaging detection. The majority of the biomacromolecules that are not captured by the nanopore 4 are still free biomacromolecules 12 in the solution.

[0078] The substrate 1 of this invention is made of silicon. Considering that in practical applications, the nanochannels need to be placed on a terahertz near-field nano-scanning platform, and the nano-scanning platform is generally a disk with a radius of about 1 cm; at the same time, considering the ease of micro-nano fabrication, the dimensions of the substrate 1 are selected as 2 cm long × 2 cm wide × 300 μm high.

[0079] The silicon nitride support layer 3 is formed by depositing a silicon nitride support layer 3 with a thickness of about 20 nanometers on the surface of the substrate 1 by plasma enhanced chemical deposition (PECVD); the inner cavity 2 is formed by etching the back side of the substrate 1 using a wet etching process.

[0080] The shape of the inner cavity 2 is not limited, but in order to make the flow of the biomacromolecule solution within the inner cavity 2 more stable during terahertz near-field detection and to facilitate the entry of biomacromolecules into the nanopore 4 under external voltage driving, the inner cavity 2 is preferably an isosceles trapezoidal groove. The bottom surface of the trapezoidal groove has a size of 1.5cm × 1.5cm, the upper surface of the trapezoidal groove has a size of 1.5cm × 2.0mm, and the height is 300 micrometers.

[0081] The bottom cover 6 and the substrate 1 are first subjected to oxygen plasma surface treatment, and then the bottom cover 6 and the substrate 1 are bonded together at room temperature. The thickness of the bottom cover 6 is 100 micrometers. The shape of the liquid inlet 7 and the liquid outlet 8 on the bottom cover 6 is not limited, and can be a round hole, a square hole, or other shapes. Preferably, the liquid inlet 7 and the liquid outlet 8 are round holes, and the diameter of the liquid inlet 7 and the liquid outlet 8 is about 100 micrometers.

[0082] in Figure 4 The diagram illustrates the detection process, in which the terahertz source 13 emits terahertz waves, the terahertz detector 14 receives terahertz near-field scattering signals, the nanoprobe 15 is used to image the biomolecular chip, and the tuning fork is connected to the nanoprobe 15. The vibration of the tuning fork 16 can drive the nanoprobe 15 to vibrate at a certain frequency.

[0083] This invention also discloses a method for manufacturing a biomolecular chip; please refer to [the relevant documentation]. Figures 1 to 4 ,include:

[0084] Etch the internal cavity onto the chip body;

[0085] At least two nanopores of uniform diameter that are connected to the inner cavity are fabricated on the chip body;

[0086] The detection window was moved to the location of nanopore 4.

[0087] Compared with the prior art, the method for manufacturing a biomolecular chip disclosed in this application can achieve the purpose of terahertz in-situ real-time imaging detection of individual biomacromolecules in solution.

[0088] In some embodiments, it also includes:

[0089] The substrate 1 is polished and ultrasonically cleaned, and a silicon nitride support layer 3 with a thickness of 20 nanometers to 30 nanometers is deposited on the surface of the substrate 1 by plasma-enhanced chemical vapor deposition to obtain the chip body.

[0090] On the back side of the substrate 1 to which the silicon nitride support layer 3 is deposited, the inner cavity is etched by a KOH wet etching process;

[0091] At least two nanopores 4 or an array of nanopores 4 with different pore sizes and connected to the inner cavity are prepared on the silicon nitride support layer 3 by ion beam. The pore size of the nanopores 4 ranges from 10 nanometers to 50 nanometers.

[0092] Graphene films with 5 or fewer layers were transferred to the nanopores 4 on the silicon nitride support layer 3 using PMMA-assisted wet transfer.

[0093] In a particularly detailed embodiment:

[0094] Microchannels are fabricated, comprising an inner cavity 2 and a silicon nitride support layer 3, with the inner cavity 3 serving as a flow channel for biomacromolecules.

[0095] Specifically, the polished and ultrasonically cleaned substrate 1 is first placed in a furnace tube. Using a PECVD system with silane, ammonia, and nitrogen as gas sources, and with controlled temperature, gas flow, and reaction time, a silicon nitride support layer 3 with a thickness of approximately 20 nm is deposited on the surface of substrate 1. Figure 2 As shown in a; then, on the back side of the substrate 1 where the silicon nitride support layer 3 is deposited, the inner cavity 2 is etched out using a KOH wet etching process, as shown in a diagram. Figure 2 As shown in b;

[0096] Fabrication of nanopores 4: Using focused helium ion beam (FIB) technology, single nanopores 4 or arrays of nanopores 4 were fabricated on the silicon nitride support layer 3 by controlling parameters such as helium ion beam current and exposure dose. The pore size ranged from 5-30 nm. Figure 2 c and Figure 3 As shown in the figure, it meets the capture requirements of different biomacromolecules.

[0097] Preparation of graphene detection window 5: A PMMA-assisted wet transfer method was used to transfer a single-layer or few-layer (2-5-layer) graphene film prepared by CVD onto a silicon nitride support layer 3 with nanopores 4 etched on it. Figure 2 As shown in d;

[0098] Microfluidic Encapsulation: Another polished and ultrasonically cleaned square substrate is selected as the bottom cover 6 of the microfluidic channel. Small holes are machined at both ends of the bottom cover 6 to serve as the inlet 7 and outlet 8 of the microfluidic channel, respectively. The unetched portion of the upper surface of the substrate and the lower surface of the bottom cover 7 are subjected to oxygen plasma surface treatment. Then, the two are bonded together at room temperature to complete the microfluidic encapsulation. Figure 2 As shown in e.

[0099] External electric field application: A gold electrode 9 is electrochemically deposited on the left edge of the graphene detection window 5. The negative terminal of the external power supply 10 is connected to the gold electrode 9, and the positive terminal of the external power supply is inserted into the microchannel through the liquid inlet 7. Figure 2 As shown in f. Due to the different electrical properties of different biological macromolecules, the positive and negative terminals of the external power source may also be connected in opposite ways.

[0100] In the description of this specification, the use of terms such as "Embodiment 1," "this embodiment," or "in one embodiment" indicates that the specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example; moreover, the specific features, structures, materials, or characteristics described may be combined in any appropriate manner in one or more embodiments or examples.

[0101] In the description of this specification, the terms "connection," "installation," "fixing," "setting," and "having" are interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances.

[0102] In the description of this specification, relational terms such as “first” and “second” are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms “comprising,” “including,” or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitation, an element defined by the phrase “comprising one…” does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0103] The above description of the embodiments is intended to enable those skilled in the art to understand and apply the technology of this invention. Those skilled in the art can easily make various modifications to these examples and apply the general principles described herein to other embodiments without creative effort. Therefore, this invention is not limited to the above embodiments. Modifications in the following situations should be within the scope of protection of this invention: ① New technical solutions implemented based on the technical solution of this invention and combined with existing common knowledge, where the technical effects of the new technical solution do not exceed the technical effects of this invention; ② Equivalent substitutions of some features of the technical solution of this invention using known technology, resulting in the same technical effects as those of this invention; ③ Extendable technical solutions based on the technical solution of this invention, where the substantive content of the extended technical solution does not exceed the technical solution of this invention; ④ Equivalent transformations made using the content of this specification and drawings, directly or indirectly applied to other related technical fields.

Claims

1. A terahertz detection biomolecule chip for use in solution environments, characterized in that, The chip includes a chip body, the chip body having an internal cavity that communicates with the outside, the surface of the chip body having at least two uniformly sized nanopores that extend into the internal cavity and are capable of accommodating a single biomacromolecule, and a detection window made of a conductive material that can be penetrated by terahertz waves is attached to the outside of the nanopores. The device that works with the biomolecular chip is equipped with a power supply, with one pole connected to the detection window and the other pole connected to the inner cavity; The chip body is connected to or etched with an oscillation circuit that controls the power supply switching and / or an intelligent control circuit that controls the current flow.

2. The terahertz detection biomolecule chip for solution environment according to claim 1, characterized in that, The pore size of the nanopores ranges from 10 nanometers to 50 nanometers.

3. The terahertz detection biomolecule chip for solution environment according to claim 1, characterized in that, The depth of the nanopores ranges from 10 nanometers to 50 nanometers.

4. A terahertz detection biomolecule chip for solution environment according to claim 1, characterized in that, The nanopores are circular, and multiple nanopores are arranged in an array.

5. A terahertz detection biomolecule chip for solution environment according to any one of claims 1 to 4, characterized in that, The inner cavity is connected to the outside through a microfluidic channel, the pore size of which ranges from 10 micrometers to 1000 micrometers.

6. A terahertz detection biomolecule chip for solution environment according to claim 5, characterized in that, The chip body includes a support layer, a substrate, and a bottom cover that are sequentially bonded together. The substrate and the support layer form the inner cavity. The support layer has the nanopores, and the bottom cover is provided with the microfluidic channels.

7. A terahertz detection biomolecule chip for solution environment according to claim 5, characterized in that, The height of the inner cavity is between 150 micrometers and 600 micrometers.

8. A method for manufacturing a biomolecular chip, used in the chip according to any one of claims 1-7, characterized in that, include: Etch the internal cavity onto the chip body; At least two nanopores of uniform diameter that communicate with the inner cavity are fabricated on the chip body; The detection window is moved to the location of the nanopore; Also includes: The substrate is polished and ultrasonically cleaned, and a silicon nitride support layer with a thickness of 20 to 30 nanometers is deposited on the substrate surface by plasma-enhanced chemical vapor deposition to obtain the chip body. On the back side of the substrate to which the silicon nitride support layer is deposited, the inner cavity is etched using a KOH wet etching process; At least two nanopores or nanopore arrays with different pore sizes and connected to the inner cavity are prepared on the silicon nitride support layer by ion beam, wherein the pore size of the nanopores ranges from 10 nanometers to 50 nanometers. A graphene film with 5 or fewer layers is transferred to the location of the nanopores on the silicon nitride support layer using a PMMA-assisted wet transfer process.

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