Single-molecule array chip based on molybdenum disulfide doping and preparation method thereof

Abstract

The present invention relates to the field of single-molecule immunoassay chips, and specifically discloses a single-molecule array chip based on molybdenum disulfide doping and a preparation method thereof. The production raw materials include a substrate chip and a molybdenum disulfide doping coating material; the molybdenum disulfide doping coating material is composed of a synergistic metal material and molybdenum disulfide according to a mass ratio of 1:0.01 to 0.8; the synergistic metal material includes ruthenium. Its advantages are: it can significantly improve the signal-to-noise ratio of the single-molecule array chip, and further improve the accuracy and reliability of this technology in single-molecule immunoanalysis.

Description

Technical Field

[0001] The present invention relates to the technical field of bio-detection chips, especially the production technology of single molecule immunoassay chips. Background Art

[0002] Single Molecule Array (SiMoA) technology is a detection method that uses single molecule counting for trace biological markers. The detection limit of this technology can reach the femtogram (fg / mL) level, which is currently the most representative single molecule immunoassay technology. The system originated from a detection system developed by the teams of David Walt and David Duffy. Its detection principle is similar to digital PCR, both of which are achieved through single molecule isolation in microreactor units combined with single molecule level signal amplification.

[0003] SiMoA technology is a single molecule immunoassay based on a microarray chip. Thousands of micron-sized micro-wells are engraved (or cast) on a millimeter-sized chip. The volume of each micro-well is about 40 fl (i.e., the single molecule array chip). Subsequently, immunocomplex magnetic beads are distributed in the micro-wells, and then the fluorescence spots are counted with the aid of a high-resolution fluorescence microscope. According to the Poisson distribution theory, the ratio of the number of wells containing both beads and fluorescent products to the total number of wells containing beads is calculated to determine the concentration of the analyte in the test sample. Its operation process is roughly as follows:

[0004] (1) Use magnetic beads with capture antibodies on the surface to capture antigens in the sample;

[0005] (2) Use detection antibodies labeled with biotin to label the captured antigens;

[0006] (3) Add streptavidin-galactosidase complex to bind to the biotin on the detection antibody;

[0007] (4) Mix the washed magnetic beads with the substrate, load them into the single molecule array chip, use a magnetic field to make the magnetic beads fall into the micro-wells that perfectly match their size, and add an oil phase to physically isolate the micro-wells;

[0008] (5) In the micro-wells containing galactosidase, the enzyme molecules catalyze the substrate to produce fluorescent products;

[0009] (6) Perform fluorescence imaging on the micro-well array, and achieve quantitative detection by comparing the number of micro-wells emitting fluorescence signals with the standard curve.

[0010] One of the main problems of the current SiMoA technology is the low sensitivity of its single-array signal, which directly affects the accuracy and reliability of detection. In practical applications, especially in single-molecule immunoassay, it is necessary to detect and analyze extremely trace amounts of molecules. However, the signal-to-noise ratio of the single-molecule array chip in the existing technology is not good, which makes it difficult to accurately extract the target signal from the background noise and limits its application breadth and accuracy in biomolecule detection. Summary of the Invention

[0011] To solve the problems existing in the prior art, the present invention provides a single-molecule array chip based on molybdenum disulfide doping and a preparation method thereof.

[0012] The technical solution adopted by the present invention is: a preparation method of a single-molecule array chip based on molybdenum disulfide doping, characterized in that: the production raw materials include a substrate chip and a molybdenum disulfide doping coating material; the molybdenum disulfide doping coating material is composed of a synergistic metal material and molybdenum disulfide in a mass ratio of 1:0.01 to 0.8; the synergistic metal material includes ruthenium.

[0013] As a further improvement of the present invention, the synergistic metal material is composed of ruthenium and palladium in a mass ratio of 1:0.3 to 1.5.

[0014] As a further improvement of the present invention, it further includes the step of forming a molybdenum disulfide doping material film on the substrate chip to obtain a coated chip. Preferably, the thickness of the molybdenum disulfide doping material film of the coated chip is 65 to 140 nm. [[ID=I5]]

[0015] As a further improvement of the present invention, the substrate chip is selected from any one of a silicon single crystal substrate chip, an FZ single crystal silicon wafer, a CZ single crystal silicon wafer, a semi-insulating single crystal silicon wafer, and an ultra-high purity single crystal silicon wafer.

[0016] The above-mentioned coated chip can be obtained by depositing the molybdenum disulfide doping coating material on the substrate chip through a thermal evaporation thin film deposition technique or by depositing the molybdenum disulfide doping coating material on the substrate chip through a magnetron sputtering coating technique. [[ID=Z2]]

[0017] If the thermal evaporation thin film deposition technique is selected for coating, the following specific steps can be carried out:

[0018] S1. The surface of the substrate chip is treated by ultrasonic cleaning or argon plasma cleaning to remove residual organic substances and oxides;

[0019] S2. Place the molybdenum disulfide doping coating material in the vacuum chamber of a thermal evaporation coating machine, place the substrate chip on a rotating coating table, control the rotation speed to be 20 to 50 rpm, and the vacuum degree to be 10 -6 ~10 -7torr, start thermal evaporation thin film deposition when the heating temperature is 500 - 600 °C; during the deposition process, the deposition rate is measured in real time by a quartz crystal microbalance, and the evaporation rate is adjusted by adjusting the heating power and the evaporation source temperature to control the deposition rate of molybdenum disulfide at the deposition rate of the synergistic metal material is controlled at

[0020] S3. Restore atmospheric pressure to release the sample by gradually increasing the ambient pressure or opening the gas inlet valve to obtain the coated chip.

[0021] The above steps can ensure the uniform and high-quality deposition of the metal thin film on the substrate surface, providing a reliable basis for the subsequent preparation of the single-molecule array chip.

[0022] As a further improvement of the present invention, it further includes the step of plasma cleaning the coated chip.

[0023] The parameters of the above plasma cleaning can be set as follows: use high-purity oxygen as the cleaning gas, set the gas pressure to 0.1 - 1 mTorr, the gas flow rate to 5 - 20 sccm, the radio frequency power to 100 - 500 W, and the cleaning time to 5 - 30 min.

[0024] The present invention also discloses a single-molecule array chip based on molybdenum disulfide doping, which is prepared by the preparation method of the single-molecule array chip based on molybdenum disulfide doping of the present invention.

[0025] The beneficial effects of the present invention are: the single-molecule array chip based on molybdenum disulfide doping prepared by the method of the present invention can significantly improve the signal-to-noise ratio of the single-molecule array chip, and further improve the accuracy and reliability of this technology in single-molecule immunoassay. Description of the Drawings

[0026] Figure 1 is the topography map of the single-molecule array chip in Example 1 under an atomic force microscope (AFM). Detailed Embodiments

[0027] The present invention will be further described below in conjunction with embodiments.

[0028] Example 1:

[0029] Prepare a single-molecule array chip according to the following steps:

[0030] (1) Prepare a molybdenum disulfide doped coating material according to the mass ratio of the synergistic metal material to molybdenum disulfide of 1:0.15, where the synergistic metal material is ruthenium;

[0031] (2) The surface of the substrate chip (silicon single crystal substrate chip, cut from a 4-inch wafer, with a thickness of 525 μm) was treated by ultrasonic cleaning to remove residual organic substances and oxides;

[0032] (3) The above-mentioned molybdenum disulfide doped coating material was placed in the vacuum chamber of a thermal evaporation coating machine, and the substrate chip was placed on a rotating coating table. The rotation speed was controlled at 45 rpm, the vacuum degree was 3×10 -7 torr, and the heating temperature was 550 °C to start thermal evaporation thin film deposition; during the deposition process, the deposition rate was measured in real time by a quartz crystal microbalance, and the evaporation rate was adjusted by adjusting the heating power and the evaporation source temperature to control the molybdenum disulfide deposition rate at The deposition rate of the synergistic metal material was controlled at When the thickness of the molybdenum disulfide doped material film on the coated chip reached 80 nm, the gas inlet valve was opened to restore atmospheric pressure to release the sample, and the coated chip was obtained.

[0033] (4) The coated chip was cleaned using a plasma cleaner (CPC-G / Gplus). The cleaning gas was high-purity oxygen (oxygen purity ≥ 99.995%), the gas pressure was set at 0.5 mTorr, the gas flow rate was 15 sccm, the radio frequency power was 400 W, and the cleaning time was 20 min. After cleaning, the atmospheric pressure was gradually restored to complete the treatment, and a single-molecule array chip was obtained.

[0034] The surface morphology of the above single-molecule array chip was observed using an atomic force microscope (AFM), and the results are as Figure 1 shown.

[0035] Example 2:

[0036] A single-molecule array chip was prepared according to the following steps:

[0037] (1) Prepare a molybdenum disulfide doped coating material according to the mass ratio of synergistic metal material to molybdenum disulfide of 1:0.3, where the synergistic metal material is ruthenium;

[0038] (2) The surface of the substrate chip (silicon single crystal substrate chip, cut from a 4-inch wafer, with a thickness of 525 μm) was treated by ultrasonic cleaning to remove residual organic substances and oxides;

[0039] (3) The above-mentioned molybdenum disulfide doped coating material was placed in the vacuum chamber of a thermal evaporation coating machine, and the substrate chip was placed on a rotating coating table. The rotation speed was controlled at 50 rpm, the vacuum degree was 2×10 -7 torr, and the heating temperature was 500 °C to start thermal evaporation thin film deposition; during the deposition process, the deposition rate was measured in real time by a quartz crystal microbalance, and the evaporation rate was adjusted by adjusting the heating power and the evaporation source temperature to control the molybdenum disulfide deposition rate at The deposition rate control of the synergistic metal material is at When the thickness of the molybdenum disulfide doped material film of the coated chip reaches 120 nm, open the gas inlet valve to restore the atmospheric pressure to release the sample, and obtain the coated chip.

[0040] (4) Use a plasma cleaner (CPC-G / Gplus) to clean the coated chip. The cleaning gas uses high-purity oxygen (oxygen purity ≥ 99.995%), set the gas pressure to 0.8 mTorr, the gas flow rate to 5 sccm, the radio frequency power to 300 W, and the cleaning time to 10 min. After the cleaning is completed, gradually restore the atmospheric pressure to complete the treatment, and obtain a single-molecule array chip.

[0041] Example 3:

[0042] This example is a control example of Example 1, and is implemented according to the same conditions and steps as Example 1, and the difference is only that: the synergistic metal material is composed of ruthenium and palladium in a mass ratio of 1:0.8 (the mass ratio of the synergistic metal material to molybdenum disulfide and the thickness of the molybdenum disulfide doped material film remain unchanged). The specific steps are as follows:

[0043] (1) Prepare the molybdenum disulfide doped coating material according to the mass ratio of the synergistic metal material to molybdenum disulfide of 1:0.15, wherein the synergistic metal material is composed of ruthenium and palladium in a mass ratio of 1:0.8;

[0044] (2) Use ultrasonic cleaning to perform surface treatment on the substrate chip (silicon single crystal substrate chip, cut from 4 inches, thickness 525 μm) to remove residual organic matter and oxides;

[0045] (3) Place the above molybdenum disulfide doped coating material in the vacuum chamber of a thermal evaporation coater, place the substrate chip on the rotating coating table, control the rotation speed to 45 rpm, the vacuum degree to 3×10 -7 torr, and start thermal evaporation thin film deposition at a heating temperature of 550 °C; during the deposition process, the deposition rate is measured in real time by a quartz crystal microbalance, and the evaporation rate is adjusted by adjusting the heating power and the evaporation source temperature to control the deposition rate of molybdenum disulfide at The deposition rate control of the synergistic metal material is at When the thickness of the molybdenum disulfide doped material film of the coated chip reaches 80 nm, open the gas inlet valve to restore the atmospheric pressure to release the sample, and obtain the coated chip.

[0046] (4) Clean the coated chip using a plasma cleaner (CPC-G / Gplus). Use high-purity oxygen (oxygen purity ≥ 99.995%) as the cleaning gas. Set the gas pressure to 0.5 mTorr, the gas flow rate to 15 sccm, the RF power to 400 W, and the cleaning time to 20 min. After cleaning, gradually restore the atmospheric pressure to complete the treatment and obtain a single-molecule array chip.

[0047] Comparative Example 1:

[0048] This comparative example is a control experiment for Example 1. It is carried out according to the same steps and conditions as Example 1, and the only difference is that: it does not include the step of coating the substrate chip. The specific implementation steps are as follows:

[0049] (1) Use ultrasonic cleaning to perform surface treatment on the substrate chip (silicon single crystal substrate chip, cut from a 4-inch piece, thickness 525 μm) to remove residual organic matter and oxides;

[0050] (2) Clean the coated chip using a plasma cleaner (CPC-G / Gplus). Use high-purity oxygen (oxygen purity ≥ 99.995%) as the cleaning gas. Set the gas pressure to 0.5 mTorr, the gas flow rate to 15 sccm, the RF power to 400 W, and the cleaning time to 20 min. After cleaning, gradually restore the atmospheric pressure to complete the treatment and obtain a single-molecule array chip.

[0051] Comparative Example 2:

[0052] This comparative example is a control experiment for Example 1. It is carried out according to the same steps and conditions as Example 1, and the only difference is that: it does not include the step of plasma cleaning the coated chip. The specific implementation steps are as follows:

[0053] (1) Prepare a molybdenum disulfide doped coating material according to the mass ratio of the synergistic metal material and molybdenum disulfide of 1:0.15, where the synergistic metal material is ruthenium;

[0054] (2) Use ultrasonic cleaning to perform surface treatment on the substrate chip (silicon single crystal substrate chip, cut from a 4-inch piece, thickness 525 μm) to remove residual organic matter and oxides;

[0055] (3) Place the above molybdenum disulfide doped coating material in the vacuum chamber of a thermal evaporation coater. Place the substrate chip on a rotating coating table, control the rotation speed to 45 rpm, the vacuum degree to 3×10 -7 torr, and the heating temperature to 550 °C to start thermal evaporation thin film deposition; during the deposition process, the deposition rate is measured in real time by a quartz crystal microbalance, and the evaporation rate is adjusted by adjusting the heating power and the evaporation source temperature to control the molybdenum disulfide deposition rate at The deposition rate control of the synergistic metal material is at When the thickness of the molybdenum disulfide doped material film of the coated chip reaches 80 nm, open the gas inlet valve to restore the atmospheric pressure to release the sample, and a single-molecule array chip is obtained.

[0056] Comparative Example 3:

[0057] This comparative example is a control experiment of Example 1, and is carried out according to the same steps and conditions as Example 1, and the only difference is that: on the premise that the total coating thickness remains unchanged, the coating material only uses the synergistic metal material (ruthenium). The specific implementation steps are as follows:

[0058] (1) Use ruthenium as the single coating material;

[0059] (2) Use ultrasonic cleaning to perform surface treatment on the substrate chip (silicon single crystal substrate chip, cut from 4 inches, thickness 525 μm) to remove residual organic substances and oxides;

[0060] (3) Place the above coating material in the vacuum chamber of a thermal evaporation coater, place the substrate chip on the rotating coating table, control the rotation speed to be 45 rpm, and the vacuum degree to be 3×10 -7 torr, and start thermal evaporation thin film deposition when the heating temperature is 550 °C; during the deposition process, the deposition rate is measured in real time by a quartz crystal microbalance, and the evaporation rate is adjusted by adjusting the heating power and the evaporation source temperature to control the deposition rate of the coating material at When the film thickness of the coated chip reaches 80 nm, open the gas inlet valve to restore the atmospheric pressure to release the sample, and a coated chip is obtained.

[0061] (4) Use a plasma cleaner (CPC-G / Gplus) to clean the coated chip, use high-purity oxygen (oxygen purity ≥ 99.995%) as the cleaning gas, set the gas pressure to 0.5 mTorr, the gas flow rate to 15 sccm, the radio frequency power to 400 W, and the cleaning time to 20 min. After the cleaning is completed, gradually restore the atmospheric pressure to complete the treatment, and a single-molecule array chip is obtained.

[0062] Comparative Example 4:

[0063] This comparative example is a control experiment of Example 1, and is carried out according to the same steps and conditions as Example 1, and the only difference is that: on the premise that the total coating thickness remains unchanged, the coating material only uses molybdenum disulfide. The specific implementation steps are as follows:

[0064] (1) Use molybdenum disulfide as the single coating material;

[0065] (2) Use ultrasonic cleaning to perform surface treatment on the substrate chip (silicon single crystal substrate chip, cut from 4 inches, thickness 525 μm) to remove residual organic substances and oxides;

[0066] (3) Place the above-mentioned molybdenum disulfide coating material in the vacuum chamber of a thermal evaporation coating machine, and place the substrate chip on a rotating coating table. Control the rotation speed to be 45 rpm and the vacuum degree to be 3×10 -7 torr, and start thermal evaporation thin film deposition at a heating temperature of 550°C. During the deposition process, the deposition rate is measured in real time by a quartz crystal microbalance, and the evaporation rate is adjusted by adjusting the heating power and the temperature of the evaporation source to control the deposition rate of molybdenum disulfide at When the thickness of the molybdenum disulfide film on the coated chip reaches 80 nm, open the gas inlet valve to restore atmospheric pressure to release the sample, and obtain the coated chip.

[0067] (4) Clean the coated chip using a plasma cleaner (CPC-G / Gplus). The cleaning gas is high-purity oxygen (oxygen purity ≥ 99.995%), set the gas pressure to 0.5 mTorr, the gas flow rate to 15 sccm, the radio frequency power to 400 W, and the cleaning time to 20 min. After cleaning, gradually restore atmospheric pressure to complete the treatment, and obtain a single-molecule array chip.

[0068] Comparative Example 5:

[0069] This comparative example is a control experiment of Example 1, which is implemented according to the same steps and conditions as Example 1, and the only difference is that: the synergistic metal material is replaced by palladium (the mass ratio of the synergistic metal material to molybdenum disulfide and the thickness of the molybdenum disulfide doping material film remain unchanged). The specific implementation steps are as follows:

[0070] (1) Prepare a molybdenum disulfide doped coating material according to the mass ratio of the synergistic metal material to molybdenum disulfide of 1:0.15, where the synergistic metal material is palladium;

[0071] (2) Use ultrasonic cleaning to perform surface treatment on the substrate chip (silicon single crystal substrate chip, cut from a 4-inch wafer, with a thickness of 525 μm) to remove residual organic substances and oxides;

[0072] (3) Place the above-mentioned molybdenum disulfide doped coating material in the vacuum chamber of a thermal evaporation coating machine, and place the substrate chip on a rotating coating table. Control the rotation speed to be 45 rpm and the vacuum degree to be 3×10 -7 torr, and start thermal evaporation thin film deposition at a heating temperature of 550°C. During the deposition process, the deposition rate is measured in real time by a quartz crystal microbalance, and the evaporation rate is adjusted by adjusting the heating power and the temperature of the evaporation source to control the deposition rate of molybdenum disulfide at Control the deposition rate of the synergistic metal material at When the thickness of the molybdenum disulfide doped material film on the coated chip reaches 80 nm, open the gas inlet valve to restore atmospheric pressure to release the sample, and obtain the coated chip.

[0073] (4) Clean the coated chip with a plasma cleaner (CPC-G / Gplus). Use high-purity oxygen (oxygen purity ≥ 99.995%) as the cleaning gas. Set the gas pressure to 0.5 mTorr, the gas flow rate to 15 sccm, the radio frequency power to 400 W, and the cleaning time to 20 min. After cleaning, gradually restore the atmospheric pressure to complete the treatment and obtain a single-molecule array chip.

[0074] Bioanalysis experiment of low-abundance substances:

[0075] Modify magnetic beads with Tau217 primary antibody, modify Tau-217 secondary antibody with NHS-Fitc, capture Tau-217 antigens with different concentrations by one-step method and sandwich method. After capture, drop the complex on the single-molecule array chip and perform statistical analysis of single-molecule signals. The specific experimental methods are as follows:

[0076] I. Experimental materials:

[0077] Single-molecule array chips prepared in Examples 1 to 3 and Comparative Examples 1 to 5;

[0078] Tau217 primary antibody: Antibody used to modify magnetic beads;

[0079] NHS-FITC: Fluorescent dye used to modify Tau217 secondary antibody;

[0080] Tau217 antigen: Target molecule for capture;

[0081] Magnetic beads: Used to modify Tau217 primary antibody;

[0082] PBS buffer: Used for dilution and washing.

[0083] II. Experimental steps:

[0084] 1. Modification of magnetic beads:

[0085] (1) Dilute Tau217 primary antibody to a final concentration of 10 μg / mL;

[0086] (2) Add Tau217 primary antibody to magnetic beads, react at 25 °C for 1 hour;

[0087] 2. Modification of Tau217 secondary antibody:

[0088] (1) Dissolve NHS-FITC in PBS buffer at a concentration of 1 mg / mL;

[0089] (2) Add NHS-FITC to the Tau217 secondary antibody to form a Tau217 secondary antibody-NHS-FITC complex. The reaction temperature is 25 °C and the reaction time is 2 hours.

[0090] 3. Capture the target molecule:

[0091] Add Tau217 antigen solutions at different concentrations (0.1 pg / mL, 0.5 pg / mL, 1 pg / mL, 10 pg / mL, 100 pg / mL, 1 ng / mL) to the modified magnetic beads using a one-step method. The reaction temperature is 4 °C and the reaction time is 12 hours.

[0092] 4. Application of the complex:

[0093] ) Drop the captured Tau217 antigen complex onto the single-molecule array chip. Use 10 μL of the complex solution for each chip.

[0094] 5. Analysis of single-molecule signal intensity:

[0095] Perform fluorescence imaging on the single-molecule array chip using a fluorescence microscope (Zeiss Axio Observer).

[0096] Detect and record the fluorescence signal using the FITC channel (excitation wavelength: 488 nm, emission wavelength: 515 nm); count the number of single-molecule signals under the capture conditions of Tau217 antigen solutions at different concentrations. The results are shown in Table 1.

[0097] Table 1 Statistical table of the number of single-molecule signals under the capture conditions of Tau217 antigen solutions at different concentrations

[0098]

[0099] It can be seen from the detection results of Examples 1 to 3 in Table 1 that the single-molecule array chip prepared by the method of the present invention can generate stable single-molecule signals under the capture conditions of Tau217 antigen solutions with concentrations ranging from 0.1 pg / mL to 1 ng / mL, and has the advantages of high detection sensitivity and stable signal intensity.

[0100] It can be seen from the comparison of the detection results of Example 1 and Comparative Example 1 in Table 1 that the single-molecule array chip of the present invention has significantly higher signal intensity compared to the ordinary single-molecule array chip without molybdenum disulfide doping coating.

[0101] It can be seen from the comparison of Example 1 and Comparative Example 2 in Table 1 that the single-molecule signal intensity can be significantly improved through the plasma cleaning step of the present invention.

[0102] From the comparison of Example 1, Comparative Example 3, and Comparative Example 4 in Table 1, it can be seen that on the premise of the same coating thickness, the single-molecule signal intensities captured by the Tau217 antigen solutions with different concentrations in Example 1 are significantly higher than those in Comparative Example 3 using only the synergistic metal (ruthenium) coating and Comparative Example 4 using only molybdenum disulfide coating. It can be seen that the synergistic metal material and molybdenum disulfide of the present invention have an obvious synergistic effect in improving the single-molecule signal intensity.

[0103] From the comparison of Example 3, Example 1, and Comparative Example 5 in Table 1, it can be seen that under the condition of the same coating thickness and the same mass ratio of the synergistic metal material and molybdenum disulfide, the single-molecule signal intensities captured by the Tau217 antigen solutions with different concentrations in Example 3 are significantly higher than those in Example 1 using only ruthenium as the synergistic metal material and Comparative Example 5 using only palladium as the synergistic metal material. It can be seen that the components of the synergistic composite metal material of the present invention have an obvious synergistic effect in improving the single-molecule signal intensity.

Claims

1. A method for preparing a single-molecule array chip doped with molybdenum disulfide, characterized in that: The production raw materials include a substrate chip and a molybdenum disulfide doped coating material; the molybdenum disulfide doped coating material is composed of a synergistic metal material and molybdenum disulfide in a mass ratio of 1:0.01 to 0.8; the synergistic metal material is composed of ruthenium and palladium in a mass ratio of 1:0.3 to 1.

5.

2. The method for preparing a single-molecule array chip based on molybdenum disulfide doping according to claim 1, characterized in that: The method also includes forming a molybdenum disulfide doping material film on the base chip to obtain a film-coated chip.

3. The method for preparing a single-molecule array chip based on molybdenum disulfide doping according to claim 2, characterized in that: The thickness of the molybdenum disulfide doping material film of the film-coated chip is 65 to 140 nm.

4. The method for preparing a single-molecule array chip based on molybdenum disulfide doping according to claim 1, characterized in that: The base chip is selected from any one of a silicon single crystal substrate base chip, an FZ single crystal silicon wafer, a CZ single crystal silicon wafer, a semi-insulating single crystal silicon wafer, and an ultra-high purity single crystal silicon wafer.

5. The method for preparing a single-molecule array chip based on molybdenum disulfide doping according to claim 2, characterized in that: The specific steps of preparing the coating chip are: depositing the molybdenum disulfide doped coating material on the base chip by thermal evaporation thin film deposition technology, thereby obtaining the coating chip.

6. The method for preparing a single molecule array chip based on molybdenum disulfide doping according to claim 5, characterized in that: The steps for preparing the coated chip are specifically as follows: S1. Surface treatment of the substrate chip is performed by ultrasonic cleaning or argon plasma cleaning to remove residual organic matter and oxides; S2, placing the molybdenum disulfide doped coating material in the vacuum chamber of the thermal evaporation coating machine, placing the substrate chip on the rotary coating table, controlling the rotation speed to 20-50 rpm, and the vacuum degree to 10 -6 ~10 -7 torr, the heating temperature is 500-600℃ to start thermal evaporation film deposition; During the deposition process, the deposition rate was measured in real time by a quartz crystal microbalance, and the evaporation rate was adjusted by adjusting the heating power and the evaporation source temperature to control the MoS2 deposition rate within The deposition rate of the enhanced metal material is controlled at S3. Restore the atmospheric pressure by gradually increasing the ambient pressure or opening the gas inlet valve to release the sample to obtain a coated chip.

7. The method for preparing a single-molecule array chip based on molybdenum disulfide doping according to claim 2, characterized in that: The method also includes a step of plasma cleaning the coated chip.

8. The method for preparing a single molecule array chip based on molybdenum disulfide doping according to claim 7, characterized in that: The plasma cleaning parameters are: using high-purity oxygen as the cleaning gas, setting the gas pressure to 0.1-1 mTorr, the gas flow rate to 5-20 sccm, the radio frequency power to 100-500 W, and the cleaning time to 5-30 min.

9. A molybdenum disulfide-doped single molecule array chip prepared by the method for preparing a molybdenum disulfide-doped single molecule array chip according to any one of claims 1 to 8.

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