Preparation method and application of micro-pit with glass as base material

Abstract

The invention belongs to the technical field of material preparation, and particularly relates to a micro-pit preparation method with glass as a base material and application of the micro-pit preparation method. The method provided by the invention mainly comprises the following steps: accelerating heavy ions by using an accelerator, then bombarding glass by using the accelerated heavy ions, after the heavy ions damage the glass, etching the glass by using an etching agent added with a corrosion inhibitor to obtain micro-pits with specific diameters and depths, and then cleaning and drying an etched sample. According to the method, the glass is impacted in an accelerated mode through heavy ions, the micro-pits are prepared, the limitation of a traditional photoetching technology is broken through, the patterns of the micro-pits are defined directly through heavy ion bombardment, a mask does not need to be used, the material cost is reduced, production interruption and extra cost caused by damage of the mask are avoided, the machining process is relatively simple, and the method is suitable for large-scale popularization and application. Multiple complex photoetching and etching steps are not needed, the production efficiency is greatly improved, the production period is shortened, and the production cost is reduced.

Description

Technical Field

[0001] This invention belongs to the field of materials preparation technology, specifically relating to a method for preparing micro-pits using glass as a substrate and its application. Background Technology

[0002] With the continuous advancement of science and technology, a variety of novel detection technologies with high sensitivity, strong specificity, and rapid diagnostic capabilities are rapidly developing and are increasingly being applied to the detection of animal diseases. Among them, single-molecule detection technology can study the structure and function of biomolecules at the single-molecule level and can be used to detect low-abundance biomarkers in complex samples. It has enormous application potential in single-molecule level imaging and tracking of interactions between low-concentration proteins, nucleic acids, and other biologically related molecules, ultrasensitive quantitative analysis, and precision medicine, and has been widely used in early disease diagnosis, exploration of potential diseases, live-cell single-virus tracing, and DNA sequencing.

[0003] Digital ELISA, as an emerging single-molecule detection technology, achieves single-molecule-level detection by dispersing single molecules into high-throughput microreactors for independent reactions and by detecting and statistically analyzing the signals from each microreactor. For example, the "Digital ELISA" technology developed by David Walt and David Duffy's team in 2010 uses labeled β-galactosidase to catalyze the substrate to generate a fluorescent signal, replacing the PCR signal amplification process in digital PCR. This allows single-molecule proteins labeled with β-galactosidase to be dispersed into high-throughput microreactors for independent reactions, thus achieving extremely high detection sensitivity. However, digital ELISA technology still faces many challenges in practical applications. Microreactor-based digital ELISA suffers from problems such as high chip fabrication difficulty and cost, and low production efficiency. Currently, the main method for fabricating microreactor detection chips is photolithography. Photolithography requires high-precision masks, complex photolithography equipment, and cumbersome experimental operations. These factors not only increase the cost of chip fabrication but also reduce its efficiency. Significant difficulties remain both technically and commercially.

[0004] Therefore, it is crucial to develop more stable fabrication methods for single-molecule detection micro-pits and novel single-molecule detection chips to improve chip fabrication efficiency, reduce chip fabrication costs, and enhance their commercial value. Summary of the Invention

[0005] To address the aforementioned challenges, this invention provides a method for fabricating micropits on a glass substrate and its applications. By using a high-energy accelerator to accelerate heavy ion bombardment of the glass substrate, the limitations of traditional photolithography techniques are overcome, significantly reducing the cost of micropit fabrication and effectively solving the problems and defects existing in the prior art.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A method for fabricating micropits using glass as a substrate includes the following steps: S1. Irradiation: Heavy ions are accelerated using an accelerator to obtain a heavy ion beam, and then the heavy ion beam is used to bombard the glass substrate to generate nuclear tracks, thereby obtaining a damaged glass substrate. S2, Etching: The damaged glass substrate obtained in step S1 is etched with an etchant to obtain a glass substrate containing micropits. S3. Cleaning and drying: Clean the glass substrate containing micro-pits obtained in step S2 with cleaning solution, and dry it in a vacuum drying oven to obtain the final product.

[0007] Preferably, in step S1, the heavy ions are accelerated to an energy of 100~1000 MeV using an accelerator, and the heavy ions are Br ions.

[0008] Preferably, the glass substrate in step S1 is a glass sheet; the thickness of the glass sheet is 0.002~1mm.

[0009] Preferably, the heavy ion beam described in step S1 is scanned into a rectangular plane using a beam scanner.

[0010] Preferably, in step S1, during the bombardment of the glass substrate with accelerated heavy ions, the glass substrate is bombarded with heavy ions in the atmosphere, and a titanium film is used for vacuum isolation. After bombarding the glass substrate, the density of the nuclear track is 1 × 10⁻⁶ per square centimeter. 4 ~5×10 7 ions.

[0011] In actual preparation, since the glass sheet is relatively thin, generally within 0.002~1mm, to avoid breakdown, this invention limits the energy of the accelerated heavy ions to 100~1000MeV. After breakdown by the accelerated Br ions, the nuclear track density of the obtained glass is 1×10⁻⁶. 4 ~5×10 7 ions / cm 2 The length of the nuclear track ranges from 0.1 to 50 micrometers.

[0012] Preferably, the etchant in step S2 is one of hydrogen fluoride solution and fluorosilicic acid solution.

[0013] Preferably, the etchant in step S2 further includes a corrosion inhibitor, which is selected from one or more of methionine and histidine.

[0014] Preferably, the etching temperature in step S2 is 20~60℃ and the etching time is 90s~600min; the diameter of the micropits on the glass substrate obtained in step S2 is 0.5~45μm and the depth of the micropits is 0.5~30μm.

[0015] In order to obtain micropits with adjustable diameter and depth, the present invention requires precise selection of etching conditions in the actual micropit preparation process: after multiple experimental screenings, hydrogen fluoride solution or fluorosilicic acid solution was finally selected as the etchant, with a concentration of 0.1~10 mol / L, and chemical etching was performed on the irradiated glass sheet at a temperature of 20~60℃ for a time of 90~600s.

[0016] However, due to the surface damage, structural changes, and enhanced chemical activity of the substrate after heavy ion bombardment, the damaged substrate is more susceptible to corrosion. If etching time and other conditions are not strictly controlled, over-corrosion can easily occur, affecting the substrate quality. Therefore, to reduce the difficulty of process control, the etchant in step S2 of this invention also includes a corrosion inhibitor, which is selected from one or more of methionine and histidine.

[0017] This invention incorporates a corrosion inhibitor during the etching of glass substrates to prevent excessive erosion of the glass sheet due to untimely monitoring or process control errors. Furthermore, by controlling the etching rate, the etching process can be monitored in real time, allowing for timely adjustments to bombardment angles and heavy ion energy. This enables the fabrication of micropits in cylindrical, conical, symmetrical biconical, and asymmetrical biconical shapes. The corrosion inhibitor prevents excessive corrosion caused by high-speed etching, thus avoiding damage to the designed shape. In particular, the biconical micropits, through their unique geometry, perfectly combine optics, fluid dynamics, and surface chemistry, achieving the goal of long-term, high signal-to-noise ratio, real-time observation of single-molecule behavior, thereby driving the birth and development of disruptive technologies such as single-molecule sequencing.

[0018] This invention, during its research on corrosion inhibitors, discovered that when one or more specific amino acids, such as methionine and histidine, are selected as the core corrosion inhibitory components, they exhibit significantly superior performance compared to traditional corrosion inhibitors. These inhibitory components can form a dense and stable protective film on the glass surface through adsorption, effectively blocking the contact between corrosive media (such as moisture and heavy ions) and the glass, while simultaneously inhibiting electrochemical corrosion reactions on the glass surface, thus significantly reducing the corrosion rate. Not only does it have high corrosion inhibition efficiency, but it can also significantly delay the process of glass rusting, peeling, or performance degradation, ultimately achieving long-term and reliable protection for the glass substrate, providing a new direction for the green and efficient application of corrosion inhibitors.

[0019] The present invention also provides a glass substrate containing micropits prepared by the above preparation method.

[0020] The present invention also provides an application of the glass substrate containing micropits in the fabrication of a single-molecule detection chip.

[0021] Compared with existing technologies, this invention achieves the fabrication of micropits by accelerating the impact of heavy ions on a glass substrate. This method overcomes the limitations of traditional photolithography and has significant advantages and beneficial effects: (1) Traditional photolithography requires the use of high-precision photomasks to define the pattern of micro-pits. The production cost of photomasks is very high and they are easily damaged. However, the present invention defines the pattern of micro-pits by heavy ion bombardment, which eliminates the need for photomasks. This not only reduces material costs but also avoids production interruptions and additional costs caused by photomask damage. (2) The present invention uses heavy ion bombardment to not only control the size of the micro-pit, but also to achieve more complex shape control. By adjusting the energy and bombardment angle of the heavy ions, micro-pits with different shapes can be prepared, such as circular, square, elliptical, etc., and even micro-pits with special structures, such as conical micro-pits, stepped micro-pits, etc., providing more possibilities for the application of micro-pits. (3) The processing of the present invention is relatively simple, without the need for multiple complex photolithography and etching steps. The micro-pits can be prepared in one heavy ion bombardment and etching process. This greatly improves production efficiency, shortens the production cycle, and reduces production costs. Attached Figure Description

[0022] Figure 1 Schematic diagrams of different shapes of micropits made from glass sheets as substrates; Figure 2 This is a schematic diagram of the micro-pits on the glass slide in Example 1. Detailed Implementation

[0023] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. These embodiments are only for better explanation of the present invention and do not limit the technical solutions of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. The reagents and equipment used in this invention are all conventional reagents and equipment in the art.

[0024] Example 1: A method for preparing micropits using glass as a substrate The specific process of the micro-pit preparation method is as follows: S1. Irradiation: Br ions are accelerated to an energy of 140 MeV using an accelerator to obtain a quasi-monoenergetic Br ion beam, which is then used to irradiate a glass slide with a thickness of 0.02 mm to generate nuclear tracks on the glass slide and obtain a damaged glass slide. S2, Etching: Using a 0.35 mol / L HF solution, chemical etching is performed on the damaged glass sheet obtained in step S1 at a temperature of 25°C for 90 seconds to obtain a glass sheet containing micropits. S3. Cleaning and drying: The glass slide containing micropits obtained in step S2 is cleaned repeatedly with clean water and deionized water. After cleaning, it is dried in a vacuum drying oven to obtain a glass slide with micropits. Figure 2 By cutting glass slides with micropits, single-molecule detection chips can be fabricated.

[0025] Example 2: A method for preparing micropits using glass as a substrate The specific process of the micro-pit preparation method is as follows: S1. Irradiation: Br ions are accelerated to an energy of 280 MeV using an accelerator to obtain a quasi-monoenergetic Br ion beam, which is then used to irradiate a glass slide with a thickness of 0.1 mm to produce nuclear tracks on the glass slide and obtain a damaged glass slide. S2. Etching: Using a 1 mol / L HF solution, chemical etching is performed on the irradiated glass slide at a temperature of 25°C for 200 seconds to obtain a glass slide containing micropits. S3. Cleaning and Drying: The glass slide containing micropits obtained in step S2 is cleaned multiple times with clean water and deionized water. After cleaning, it is dried in a vacuum drying oven to obtain a glass slide with micropits. The glass slide with micropits is then cut to make a chip for single-molecule detection.

[0026] Example 3: A method for preparing micropits using glass as a substrate The specific process of the micro-pit preparation method is as follows: S1. Irradiation: Br ions are accelerated to an energy of 560 MeV using an accelerator to obtain a quasi-monoenergetic Br ion beam, which is then used to irradiate a glass slide with a thickness of 0.35 mm to generate nuclear tracks on the glass slide and obtain a damaged glass slide. S2. Etching: Using a 4 mol / L HF solution, chemical etching is performed on the irradiated glass slide at a temperature of 50°C for 360 seconds to obtain a glass slide containing micropits. S3. Cleaning and Drying: The glass slide containing micropits obtained in step S2 is cleaned multiple times with clean water and deionized water. After cleaning, it is dried in a vacuum drying oven to obtain a glass slide with micropits. The glass slide with micropits is then cut to make a chip for single-molecule detection.

[0027] Example 4: A method for preparing micropits using glass as a substrate The specific process of the micro-pit preparation method is as follows: S1. Irradiation: Br ions are accelerated to an energy of 980 MeV using an accelerator to obtain a quasi-monoenergetic Br ion beam, which is then used to irradiate a glass slide with a thickness of 1 mm to produce nuclear tracks on the glass slide, thus obtaining a damaged glass slide. S2. Etching: Using a 10 mol / L HF solution, chemical etching is performed on the irradiated glass slide at a temperature of 50°C for 500 seconds to obtain a glass slide containing micropits. S3. Cleaning and Drying: The glass slide containing micropits obtained in step S2 is cleaned multiple times with clean water and deionized water. After cleaning, it is dried in a vacuum drying oven to obtain a glass slide with micropits. The glass slide with micropits is then cut to make a chip for single-molecule detection.

[0028] Example 5: A method for preparing micropits using glass as a substrate The specific process of the micro-pit preparation method is as follows: S1. Irradiation: Br ions are accelerated to an energy of 980 MeV using an accelerator to obtain a quasi-monoenergetic Br ion beam, which is then used to irradiate a glass slide with a thickness of 1 mm to produce nuclear tracks on the glass slide, thus obtaining a damaged glass slide. S2. Etching: Using a 10 mol / L HF solution, a certain weight of methionine is added to the solution until the final concentration of methionine is 0.7 g / L. The irradiated glass slide is chemically etched at a temperature of 50°C for 600 s to obtain a glass slide containing micropits. S3. Cleaning and Drying: The glass slide containing micropits obtained in step S2 is cleaned multiple times with clean water and deionized water. After cleaning, it is dried in a vacuum drying oven to obtain a glass slide with micropits. The glass slide with micropits is then cut to make a chip for single-molecule detection.

[0029] Comparative Example 1: A method for preparing micropits The method for preparing the micro-pits is similar to that in Example 5; The difference from Example 5 is that in Comparative Example 1, methionine was replaced with an equal amount of cystine during etching, and the final concentration of cystine was 0.1 g / L.

[0030] Comparative Example 2: A method for preparing micropits The method for preparing the micro-pits is similar to that in Example 5; The difference from Example 5 is that Comparative Example 2 uses bismuth ions as heavy ions.

[0031] Comparison of Micropit Parameters in Experiment Example 1 1. Experimental sample: Glass slides with micro-pits prepared in Examples 1-5.

[0032] 2. Experimental method: During the experiment, the density and length of nuclear tracks generated on the glass slide during irradiation were recorded in real time, and the opening diameter and depth of individual micropits were measured after cleaning and drying.

[0033] 3. Experimental results: The specific experimental results are shown in Table 1.

[0034] Table 1. Parameters related to micropits on glass slides as substrates

[0035] As shown in Table 1 above, this invention obtains nuclear tracks of different densities and lengths by adjusting the heavy ion beam energy and irradiation time. The change in heavy ion beam energy has a significant impact on the density of the nuclear tracks, ranging from 10... 4 ~10 7 Furthermore, during the etching process, etching temperature and time directly affect the size and depth of the micropores. As temperature and time increase, the diameter and depth of the micropores gradually increase. However, the addition of an etching inhibitor alters the diameter and depth of the micropores to varying degrees. This demonstrates that by changing the etching temperature and time, micropores of different sizes can be prepared, effectively improving the preparation efficiency and laying the foundation for their application in single-molecule detection.

[0036] Experiment Example 2: Comparison of Micro-pit Shapes 1. Experimental samples: Micropits prepared in Example 5 and micropits prepared in Comparative Examples 1-2; 2. Experimental method: The sample surface was scanned with a focused electron beam to observe the shape of different micro-pits; 3. Experimental Results: Taking Example 5 as an example, the possible effects of controlling heavy ion energy and angle are as follows: By adjusting the bombardment angle, heavy ion energy, etc., a cylindrical hole can be obtained ( Figure 1 a) Conical hole ( Figure 1 (b) Symmetrical double conical hole ( Figure 1 c) Asymmetric double conical hole ( Figure 1 In addition to the micropits of shapes such as d), unconventional shapes such as hemispherical, ellipsoidal, or teardrop-shaped micropits can also be obtained. These micropits can be further used for single-molecule detection. In contrast, in Comparative Example 1, due to the change in the type of corrosion inhibitor, the etching rate control effect decreased during etching, making it impossible to obtain the aforementioned micropit shapes, and over-etching easily occurred, leading to penetration. In Comparative Example 2, the glass slide was irradiated with a bismuth ion beam. Since bismuth is a highly inert metallic element, it hardly participates in chemical reactions during ion beam processing, and its material removal almost entirely relies on physical sputtering. Therefore, the etching selectivity is very poor, making it difficult to achieve precise stop control and control the shape of the micropits.

Claims

1. A method for preparing micropits using glass as a substrate, characterized in that, The method comprises the following steps: S1, irradiation: accelerating heavy ions by an accelerator to obtain a heavy ion beam, and then irradiating a glass substrate by the heavy ion beam to generate nuclear tracks and obtain a damaged glass substrate; S2, etching: etching the damaged glass substrate obtained in step S1 by an etchant to obtain a glass substrate containing micro-pits; S3, cleaning and drying: cleaning the glass substrate containing micro-pits obtained in step S2 by a cleaning solution, and drying in a vacuum drying oven to obtain the glass substrate containing micro-pits.

2. The micro-pit producing method according to claim 1, wherein In step S1, the heavy ions are accelerated to an energy of 100-1000 MeV by the accelerator, and the heavy ions are Br ions.

3. The micro-pit producing method according to claim 1, wherein In step S1, the glass substrate is a glass sheet, and the thickness of the glass sheet is 0.002-1 mm.

4. The micro-pit producing method according to claim 1, wherein In step S1, the heavy ion beam is scanned into a rectangular plane by a beam scanner.

5. The micro-pit producing method according to claim 1, wherein Step S1, in the process of bombarding the glass substrate with accelerated heavy ions, the heavy ions are used to bombard the glass substrate under atmosphere, and a titanium film is used for vacuum isolation. After the glass substrate is bombarded, the density of nuclear tracks is 1 x 10 4 5 x 10 7 ions per square centimeter. In the process of bombarding the glass substrate with accelerated heavy ions, the heavy ions are used to bombard the glass substrate under atmosphere, and a titanium film is used for vacuum isolation. After the glass substrate is bombarded, the density of nuclear tracks is 1 x 10 4 5 x 10 7 ions per square centimeter.

6. The micro-pit producing method according to claim 1, wherein In step S2, the etchant is one of hydrogen fluoride solution and fluorosilicic acid solution.

7. The micro-pit producing method according to claim 6, wherein In step S2, the etchant further comprises a corrosion inhibitor, and the corrosion inhibitor is one or more selected from methionine and histidine.

8. The micro-pit producing method according to Claim 1, wherein In step S2, the etching temperature is 20-60℃, and the etching time is 90 s-600 min; the diameter of the micro-pits on the glass substrate obtained in step S2 is 0.5-45 μm, and the depth of the micro-pits is 0.5-30 μm.

9. A glass substrate containing micro-pits prepared by the method of any one of claims 1-8.

10. Use of the glass substrate containing micro-pits of claim 9 in the preparation of a single molecule detection chip.

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