Detection chip and preparation method thereof

Abstract

The invention provides a detection chip and a preparation method thereof. The detection chip comprises a substrate material layer; the feature array comprises a plurality of feature units distributed on the substrate material layer; the feature array protection structure is formed on the substrate material layer, the feature array is distributed in the feature array protection structure, and the feature array protection structure at least comprises an adsorption material used for adsorbing a biological sample. The excitation power enhanced by the detection chip is at least improved by multiple orders of magnitude compared with the incident laser power, the enhancement proportion can be further improved along with characteristic array design optimization and process improvement, and therefore the excitation power received by the fluorescent dye is improved under the condition that the laser power is not changed, and the fluorescence intensity of the fluorescent dye is improved. Compared with the prior art, the method has the advantages that higher fluorescence signal intensity is generated, so that the exposure time is effectively shortened and the sequencing flux is improved under the condition that the imaging quality and the power of the laser are not changed.

Description

Detection chip and preparation method thereof Technical Field

[0001] The present application relates to the technical field of biological sample testing, and in particular to a biological sample detection chip and a preparation method thereof. Background Art

[0002] Currently, the fluorescence signal intensity of biological sample detection chips, such as sequencing chips, is proportional to the laser power, the quantum yield of the fluorescent dye, and the exposure time. Given a specific fluorescent dye model, improvements in sequencing throughput are typically achieved by reducing exposure time, but the imaging quality, that is, the fluorescence signal intensity, must remain constant. Therefore, the laser power needs to be increased, which brings with it a number of unfavorable factors: first, the cost of high-power lasers increases exponentially; second, optical components with higher damage thresholds need to be customized, increasing processing difficulty and cost; third, thermal effects, when reaching a certain level, can affect the imaging quality of the objective lens, leading to a decrease in sequencing accuracy; fourth, laser background noise increases, and the signal-to-noise ratio decreases, requiring the use of cameras with higher quantum efficiency, which also increases costs. These numerous unfavorable factors restrict the improvement of sequencing system throughput and the reduction of hardware costs, so it is necessary to seek breakthroughs from other angles.

[0003] Figure 2 shows the basic structure of a sequencing chip. The substrate is silicon (Si), with DNA (deoxyribonucleic acid) attached to the SiO2 layer. When laser light shines on the sequencing chip, some of the laser light passes through the DNA and strikes the substrate, while only a portion is reflected back and strikes the DNA again. Therefore, the laser power actually absorbed by the fluorescent dye on the DNA is less than the incident laser power. Some sequencing chips use a glass substrate, which has a high transmittance. Light that passes through the DNA is essentially not reflected back again, resulting in lower laser power utilization.

[0004] Currently, some technologies increase laser power utilization to a certain extent by adding a reflective coating to the substrate, but they cannot effectively achieve further enhancement of laser power. With the development of sequencing technology, sequencing costs are constantly being reduced, so there is a need to increase sequencing throughput and reduce hardware costs. For traditional sequencing chips, increasing sequencing throughput and reducing hardware costs are not mutually exclusive. Increasing sequencing throughput means requiring higher-power lasers, optical components with higher loss thresholds, and more sensitive detectors, which increases hardware costs exponentially.

[0005] Summary of the Invention

[0006] The main purpose of this application is to provide a sequencing chip and a preparation method thereof to improve the above-mentioned defects existing in the prior art.

[0007] This application solves the above technical problems through the following technical solutions:

[0008] As one aspect of the present application, a detection chip is provided, comprising:

[0009] a base material layer;

[0010] a feature array comprising a plurality of feature units distributed on the base material layer; and

[0011] A feature array protection structure is formed on the base material layer, and the feature array is distributed within the feature array protection structure, wherein the feature array protection structure at least includes an adsorption material for adsorbing biological samples.

[0012] As an optional implementation, the plurality of characteristic units are regularly distributed on the base material layer.

[0013] As an optional embodiment, the plurality of characteristic units are periodically and regularly distributed on the base material layer.

[0014] As an optional embodiment, the detection chip further includes a sidewall material layer formed on the feature array protection structure;

[0015] The sidewall material layer includes a plurality of sidewall materials. The sample holes formed on the feature array protection structure by adjacent sidewall materials correspond one to one with the positions of the feature units along the extension direction of the feature units. The sample holes are used to accommodate the biological samples.

[0016] As an optional embodiment, the feature array protection structure includes a feature array protection layer and an adsorption material layer formed by the adsorption material;

[0017] The feature array protection layer is formed on the base material layer, the feature array is distributed in the feature array protection layer, the adsorption material layer is formed on the feature array protection layer, and the sidewall material layer is formed on the adsorption material layer.

[0018] As an optional embodiment, the feature array protection layer includes a first feature array protection layer and a second feature array protection layer;

[0019] The feature array is distributed in the first feature array protection layer, and the second feature array protection layer is formed on the first feature array protection layer and contacts the lower surface of the adsorption material layer.

[0020] As an optional embodiment, the thickness of the feature array protection layer ranges from 5 nm to 200 nm; and / or,

[0021] The thickness of the adsorption material layer is in the range of 5 nm to 200 nm; and / or,

[0022] The thickness of the sidewall material layer ranges from 10 nm to 1000 nm.

[0023] As an optional embodiment, the material of the feature array protection layer includes any one or more of SiO2 (silicon dioxide), Si3N4 (silicon nitride), and TiO2 (titanium dioxide); and / or,

[0024] The material of the sidewall material layer includes any one or more of SiO2 and Si3N4.

[0025] As an optional embodiment, the feature array protection structure includes an adsorption material layer formed by the adsorption material;

[0026] The adsorption material layer is formed on the base material layer, the feature array is distributed in the adsorption material layer, and the sidewall material layer is formed on the adsorption material layer.

[0027] As an optional embodiment, the adsorption material layer includes a first adsorption material layer and a second adsorption material layer;

[0028] The feature array is distributed in the first adsorption material layer, and the second adsorption material layer is formed on the first adsorption material layer and contacts the lower surface of the sidewall material layer.

[0029] As an optional embodiment, the feature array protection structure includes a feature array protection layer and a sample adsorption layer formed by the adsorption material;

[0030] The feature array protection layer is formed on the base material layer, the feature array is distributed in the feature array protection layer, and the sample adsorption layer is formed on the feature array protection layer;

[0031] The sample adsorption layer includes a plurality of passivation regions and a plurality of amination regions. The amination regions correspond one-to-one to the positions of the characteristic units along the extension direction of the characteristic units. The amination regions are used to adsorb the biological sample.

[0032] As an optional embodiment, the feature array protection layer includes a first feature array protection layer and a second feature array protection layer;

[0033] The feature array is distributed in the first feature array protection layer, and the second feature array protection layer is formed on the first feature array protection layer and contacts the lower surface of the sample adsorption layer.

[0034] As an optional embodiment, the material of the base material layer includes any one or more of silicon, quartz or glass.

[0035] As an optional embodiment, the adsorption material includes any one or more of TiN (titanium nitride), TiO2, Ag (silver), ZrO2 (zirconium dioxide), and ZnO (zinc oxide).

[0036] As an optional embodiment, the feature array includes a metal array, and the feature unit includes a metal material.

[0037] As an optional embodiment, the metal material includes any one or more of Ag, Au (gold), Cu (copper), and Al (aluminum).

[0038] As an optional embodiment, the plurality of characteristic units are periodically distributed on the base material layer in any one or more forms of polygon, circle, or ellipse; and / or,

[0039] The structure of the characteristic unit includes a cylinder or a sphere; and / or,

[0040] The cross-sectional shape of the characteristic unit includes polygon, circle or ellipse.

[0041] As an optional embodiment, the thickness of the characteristic unit ranges from 20 nm to 300 nm; and / or,

[0042] The cross-sectional size of the characteristic unit ranges from 20 nm to 300 nm; and / or,

[0043] The distance between the centers of two adjacent characteristic units ranges from 100 nm to 1000 nm.

[0044] As an optional embodiment, the top surface of the feature array protection structure is higher than the top surface of any of the feature units.

[0045] As another aspect of the present application, a method for preparing a detection chip is provided, and the detection chip as described above is prepared by the preparation method;

[0046] The preparation method comprises:

[0047] providing a base material layer;

[0048] forming a feature array on the base material layer, wherein the feature array includes a plurality of feature units distributed on the base material layer;

[0049] A feature array protection structure is formed on the base material layer so that the feature array is distributed within the feature array protection structure, wherein the feature array protection structure at least includes an adsorption material for adsorbing biological samples.

[0050] Based on the content of this application, those skilled in the art can understand other aspects of the content of this application.

[0051] The positive progress of this application is:

[0052] This application provides a novel biological sample detection chip based on localized surface plasmon enhancement and its preparation method. After the detection chip is enhanced, the excitation power is increased by at least 6 to 10 orders of magnitude compared to the incident laser power. With the optimization of the feature array design and process improvements, the enhancement ratio can be further increased, thereby increasing the excitation power received by the fluorescent dye without changing the laser power, generating a stronger fluorescence signal intensity. Therefore, it is possible to effectively reduce the exposure time and increase the sequencing throughput while ensuring that the imaging quality and laser power remain unchanged. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The features and advantages of the present invention will be better understood after reading the detailed description of the embodiments of the present invention in conjunction with the following drawings. In the drawings, components are not necessarily drawn to scale, and components with similar related properties or features may have the same or similar reference numerals.

[0054] FIG1 is a schematic diagram of a partial structure of a detection chip according to an embodiment of the present application.

[0055] FIG2 is a schematic diagram of the basic structure of an existing sequencing chip.

[0056] FIG3 is a schematic diagram of a partial structure of a detection chip of a first structure according to an embodiment of the present application.

[0057] FIG4 is a schematic diagram showing the three-dimensional structure of a cylindrical characteristic unit.

[0058] FIG5 is a schematic diagram showing the three-dimensional structure of a triangular prism-shaped characteristic unit.

[0059] FIG6 is a schematic diagram showing the three-dimensional structure of a cubic characteristic unit.

[0060] FIG. 7 is a schematic diagram showing the three-dimensional structure of a spherical characteristic unit.

[0061] FIG8 is a schematic structural diagram showing the periodic distribution of quadrilaterals of cylindrical characteristic units.

[0062] FIG9 is a schematic structural diagram of the minimum period unit of the feature array in FIG8 .

[0063] FIG10 is a schematic structural diagram showing the hexagonal periodic distribution of cylindrical characteristic units.

[0064] FIG11 is a schematic structural diagram of the minimum period unit of the feature array in FIG10 .

[0065] FIG12 is a schematic structural diagram showing the circular periodic distribution of cylindrical characteristic units.

[0066] FIG13 is a schematic structural diagram of the minimum period unit of the feature array in FIG12 .

[0067] FIG. 14 is a schematic diagram illustrating the electric field distribution formed on the surface of a feature array.

[0068] FIG. 15 is a schematic diagram illustrating the electric field distribution formed between two cylindrical feature units.

[0069] FIG16 is a schematic diagram showing the optical path of an imaging device of a detection chip according to an embodiment of the present application.

[0070] FIG17 is a schematic diagram of a partial structure of a detection chip of a second structure according to an embodiment of the present application.

[0071] FIG18 is a schematic diagram of a partial structure of a detection chip with a third structure according to an embodiment of the present application.

[0072] FIG19 is a schematic flow chart of a method for preparing a detection chip according to another embodiment of the present application.

[0073] FIG20 is a schematic diagram of a processing process of a method for preparing a detection chip according to another embodiment of the present application. DETAILED DESCRIPTION

[0074] The present application is further described below by way of examples, but the present application is not limited to the scope of the examples.

[0075] It should be noted that references in the specification to "one embodiment," "an alternative embodiment," "another embodiment," etc. indicate that the described embodiment may include a particular feature, structure, or characteristic, but every embodiment may not necessarily include that particular feature, structure, or characteristic. Moreover, such phrases do not necessarily refer to the same embodiment. In addition, when a particular feature, structure, or characteristic is described in conjunction with an embodiment, whether or not explicitly described, it is within the knowledge of those skilled in the relevant art to implement such feature, structure, or characteristic in conjunction with other embodiments.

[0076] In the description of the contents of this application, it should be understood that the terms "center", "lateral", "up", "down", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the contents of this application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the contents of this application. In addition, the terms "first" and "second" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, features defined as "first" and "second" may explicitly or implicitly include one or more of such features. In the description of the contents of this application, unless otherwise specified, "multiple" means two or more. In addition, the term "including" and any variations thereof are intended to cover non-exclusive inclusions.

[0077] In the description of the present application, it should be noted that, unless otherwise expressly specified or limited, the terms "installed," "connected," and "connected" should be understood in a broad sense. For example, they can refer to fixed connections, detachable connections, or integral connections; they can refer to mechanical connections or electrical connections; they can refer to direct connections or indirect connections through an intermediate medium; and they can refer to internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present application in specific contexts.

[0078] The terms used herein are intended only to describe specific embodiments and are not intended to limit exemplary embodiments. Unless the context clearly indicates otherwise, the singular forms "a", "an", "an item" used herein are also intended to include the plural. It should also be understood that the terms "comprise" and / or "include" used herein specify the presence of stated features, integers, steps, operations, units and / or components, and do not preclude the presence or addition of one or more other features, integers, steps, operations, units, components and / or combinations thereof.

[0079] In order to overcome the above-mentioned defects that currently exist, as shown in Figure 1, this embodiment provides a detection chip, which mainly includes: a base material layer 11; a feature array 12, including a plurality of feature units 121 distributed on the base material layer 11; and a feature array protection structure 13, formed on the base material layer 11, and the feature array 12 is distributed in the feature array protection structure 13, wherein the feature array protection structure 13 at least includes an adsorption material 1311 for adsorbing biological samples.

[0080] In this embodiment, the detection chip is preferably a sequencing chip, but is not limited to sequencing chips. Other biological or chemical detection chips can also be used, and adjustments or selections can be made based on actual or potential needs. In addition to being used in the sequencing field, the detection chip can also be applied to various other fields such as protein analysis, single-cell analysis, and drug screening. While achieving the functions described above, adjustments or selections can be made based on actual or potential needs.

[0081] In this embodiment, the biological sample can be, for example, DNA beads, quantum dots, fluorescent nanospheres, etc. in a sequencer. Of course, this embodiment does not specifically limit the type of sample, and corresponding settings and adjustments can be made according to actual needs, actual scenarios, or possible needs and scenarios.

[0082] In this embodiment, the top surface of the feature array protection structure 13 is at least higher than the top surface of any feature unit 121 , so that the feature array protection structure 13 can effectively protect each feature unit 121 .

[0083] In this embodiment, a novel biological sample detection chip based on localized surface plasmon enhancement is provided. After the detection chip is enhanced, the excitation power is increased by at least 6 to 10 orders of magnitude compared to the incident laser power. With the optimization of the feature array design and process improvements, the enhancement ratio can be further increased, thereby increasing the excitation power received by the fluorescent dye without changing the laser power, generating a stronger fluorescence signal intensity. Therefore, the exposure time can be effectively reduced and the sequencing throughput can be improved while ensuring that the imaging quality and laser power remain unchanged.

[0084] In this embodiment, a nano-feature array structure is used on the surface of the detection chip, and a local surface plasma effect is generated when the laser is incident, thereby obtaining an excitation power enhancement effect. As shown in Figure 16, Figure 16 shows a schematic diagram of the optical path when the imaging device of the detection chip emits a laser. Specifically, the laser 21 is converted into parallel light by the collimating lens 22 and irradiated onto the excitation filter 23. The filtered laser is irradiated onto the dichroic mirror 24. Thereafter, the laser is reflected to the rear end face of the objective lens 25. The objective lens 25 focuses the laser illumination onto the chip assembly 26 containing the detection chip provided in this embodiment. The nano-feature array on the detection chip generates local surface plasma, and the fluorescent dye of the DNA on the chip, the generated fluorescence signal is collected by the objective lens 25 and then irradiated onto the emission filter 27 through the dichroic mirror 24, and finally focused by the lens 28 onto the detector 29 to obtain a sequencing image.

[0085] Among them, local surface plasmon enhancement means that when the laser is irradiated on the nano-feature unit, when the laser frequency is the same as the vibration frequency of the nano-feature unit, the nano-feature unit will have a strong absorption effect on the light, and a local surface plasmon resonance phenomenon will occur, forming an electric field distribution on the surface of the nano-feature unit. As shown in Figure 14, the electric field intensity at the sharp corner of the nano-feature unit increases by 3 to 4 orders of magnitude compared with the incident light intensity. For some specially designed nano-feature array structures, such as between two cylinders, as shown in Figure 15, the electric field intensity will be enhanced to 6 to 10 orders of magnitude.

[0086] The resonant frequency of a nano-feature unit is closely related to the electron density, effective electron mass, shape, and size of the charge distribution. Therefore, the local surface plasmon resonance effect can be adjusted by adjusting the material, size, shape, and surrounding dielectric material of the nano-feature unit. The structural parameters are simulated and calculated using the commonly used optical simulation software FDTD (Finite Difference Time Domain). The corresponding detection chip is then processed and fed back to the simulation model after actual measurement. After the simulation model is corrected, the optimal structural parameters are further optimized to achieve the maximum power enhancement effect.

[0087] Specifically, as an optional embodiment, as shown in Figure 3, the detection chip as the first structure mainly includes: a base material layer 11, a feature array including a number of feature units 121 regularly distributed on the base material layer 11, a feature array protection structure and a sidewall material layer formed on the feature array protection structure.

[0088] The feature array protection structure includes a feature array protection layer 132 and an adsorption material layer 131 formed of an adsorption material. The feature array protection layer 132 is formed on the base material layer 11, with the feature array distributed within the feature array protection layer 132. The adsorption material layer 131 is formed on the feature array protection layer 132, and the sidewall material layer is formed on the adsorption material layer 131. In this embodiment, the feature array protection layer 132 can be further divided into a first feature array protection layer and a second feature array protection layer (not specifically distinguished in the figure). The feature array is distributed within the first feature array protection layer (whose thickness can be similar to that of the feature cells 121). The second feature array protection layer is formed on the first feature array protection layer and contacts the lower surface of the adsorption material layer 131. The sidewall material layer includes a plurality of sidewall materials 14. Sample wells 15 formed in the adsorption material layer 131 by adjacent sidewall materials 14 correspond one-to-one with the positions of the feature cells 121 along the extension direction of the feature cells 121. The sample wells 15 are used to accommodate biological samples.

[0089] Among them, the materials of the first characteristic array protective layer and the second characteristic array protective layer can be the same or different, the materials of the adsorption material layer 131 can be the same or different from the materials of the first characteristic array protective layer and the second characteristic array protective layer, and the materials of the sidewall material layer can be the same or different from the materials of the first characteristic array protective layer and the second characteristic array protective layer. Therefore, corresponding selections and adjustments can be made according to actual needs or possible needs.

[0090] As an optional embodiment, the material of the base material layer 11 includes any one or more of silicon, quartz or glass, but is not limited thereto, and can be adjusted and selected accordingly according to actual needs or possible needs.

[0091] As an optional implementation, the feature unit 121 may be made of metal material, that is, the feature array may be a metal array, but it is not limited thereto and may be selected and adjusted accordingly according to actual needs or possible needs.

[0092] The metal material includes any one or more of Ag, Au, Cu, and Al, but is not limited thereto, and can be adjusted and selected accordingly according to actual needs or possible needs.

[0093] As an optional embodiment, several feature units 121 are distributed on the base material layer 11 in any one or more forms of polygonal, circular, and elliptical shapes. The structure of the feature unit 121 includes a cylinder or a sphere, and the cross-sectional shape of the feature unit 121 includes a polygonal, circular, or elliptical shape, but is not limited to this. They can be adjusted and selected accordingly according to actual needs or possible needs.

[0094] Specifically, in terms of the structure and shape of the characteristic unit: as shown in Figure 4, the characteristic unit can be cylindrical; as shown in Figure 5, the characteristic unit can be triangular prism-shaped; as shown in Figure 6, the characteristic unit can be cube-shaped; as shown in Figure 7, the characteristic unit can be spherical.

[0095] As a preferred embodiment, a plurality of characteristic units 121 are periodically and regularly distributed on the base material layer 11 .

[0096] In terms of the periodic regular distribution of the characteristic units: as shown in Figures 8 and 9, the characteristic array can be a quadrilateral periodic distribution; as shown in Figures 10 and 11, the characteristic array can be a hexagonal periodic distribution; as shown in Figures 12 and 13, the characteristic array can be a circular periodic distribution.

[0097] In this embodiment, the thickness and cross-sectional dimensions (such as diameter, etc.) of the characteristic unit will affect the local surface plasmon effect and be limited by the nano-processing capability. The thickness of the characteristic unit can range from 20nm to 300nm, preferably 60nm in thickness; the cross-sectional dimensions of the characteristic unit can range from 20nm to 300nm, preferably 100nm in cross-sectional dimension; the period, that is, the interval between the centers of two adjacent characteristic units, can be determined according to the diffraction limit of the optical system. The interval between the centers of two adjacent characteristic units ranges from 100nm to 1000nm, preferably 700nm.

[0098] As an optional embodiment, the material of the feature array protection layer 132 includes any one or more of SiO2, Si3N4, and TiO2, but is not limited thereto, and can be adjusted and selected accordingly according to actual needs or possible needs.

[0099] In this embodiment, the thickness of the feature array protection layer 132 ranges from 5 nm to 200 nm, preferably 10 nm. If the thickness is less than 5 nm, it is difficult to achieve the effect of protecting the metal, and if the thickness is greater than 200 nm, the local surface plasmon energy is low.

[0100] As an optional embodiment, the adsorption material includes any one or more of TiN, TiO2, Ag, ZrO2, and ZnO, but is not limited thereto, and can be adjusted and selected accordingly according to actual needs or possible needs.

[0101] In this embodiment, the thickness of the adsorption material layer 131 ranges from 5 nm to 200 nm, preferably 10 nm. If the thickness is less than 5 nm, it is difficult to achieve the metal protection effect, and if the thickness is greater than 200 nm, the local surface plasmon energy is low.

[0102] As an optional embodiment, the material of the sidewall material layer includes any one or more of SiO2, Si3N4, but is not limited thereto, and can be adjusted and selected accordingly according to actual needs or possible needs.

[0103] In this embodiment, the thickness of the sidewall material layer ranges from 10 nm to 1000 nm, preferably 100 nm. The thickness of the sidewall material layer is determined according to the size of the biological sample (eg, the diameter of the DNA nanosphere).

[0104] In this embodiment, the parameters of each layer (material, thickness, diameter, pitch (the distance between the centers of two adjacent characteristic units), shape, periodic distribution, etc.) can be simulated and confirmed using the optical simulation software FDTD according to actual usage conditions (laser wavelength, incident angle, spot uniformity, fluorescent dye, etc.), and the actual test results of the processed detection chip are fed back to the simulation software. After further optimization and simulation, the optimal detection chip parameters are obtained to achieve maximum excitation power enhancement.

[0105] As a specific embodiment, according to the actual usage scenario of this embodiment (532nm & 640nm laser wavelength, 0 degree incidence, imaging area illumination uniformity greater than 95%, fluorescent dyes are AF532 & Cy5 (fluorescent dye type)), the obtained better detection chip has the following structures: 8-inch silicon-based wafer, nano-array Ag cylinder (60nm height, 130nm diameter, 700nm pitch, square periodic distribution), SiO2 feature array protection layer (filling the gap between the Ag cylinders and 10nm higher than the top of the Ag cylinder), TiO2 adsorption material layer (DNA sample can be adsorbed on TiO2, thickness 10nm), SiO2 sidewall material layer (sidewall material height 100nm, distribution period is similar to the feature array distribution period).

[0106] As another optional embodiment, as shown in Figure 17, the detection chip as the second structure mainly includes: a base material layer 11, a feature array including a plurality of feature units 121 distributed on the base material layer 11, a feature array protection structure and a sidewall material layer formed on the feature array protection structure.

[0107] The feature array protection structure includes an adsorption material layer 131 formed by adsorption material. The adsorption material layer 131 is formed on the base material layer 11 . The feature array is distributed in the adsorption material layer 131 . The sidewall material layer is formed on the adsorption material layer 131 .

[0108] In this embodiment, the adsorption material layer 131 can be further divided into a first adsorption material layer and a second adsorption material layer (not specifically distinguished in the figure), the feature array is distributed in the first adsorption material layer (whose thickness can be similar to the thickness of the feature unit 121), and the second adsorption material layer is formed on the first adsorption material layer and contacts the lower surface of the sidewall material layer.

[0109] The sidewall material layer includes a plurality of sidewall materials 14 . The sample holes 15 formed on the adsorption material layer 131 by adjacent sidewall materials 14 correspond one-to-one to the positions of the feature units 121 along the extending direction of the feature units 121 . The sample holes 15 are used to accommodate biological samples.

[0110] Among them, the materials of the first adsorption material layer and the second adsorption material layer can be the same or different, and the materials of the side wall material layer can be the same or different from those of the first adsorption material layer and the second adsorption material layer, respectively. Therefore, corresponding selection and adjustment can be made according to actual needs or possible needs.

[0111] In this embodiment, the detection chip of the second structure is similar to the detection chip of the first structure in terms of the materials, sizes, distribution and shape of the feature arrays of each layer. Therefore, the specific parameters of the detection chip of the first structure can be referred to, and will not be described one by one.

[0112] In this embodiment, in the detection chip of the second structure, the feature array protection layer and the adsorption material layer are merged, and only the adsorption material layer is used, thereby achieving the effects of sample adsorption and protection of the feature array at the same time, thereby simplifying the structure. In addition, since the local surface plasma intensity is related to the distance between the feature arrays, the smaller the distance, the better the enhancement effect. Therefore, by merging the feature array protection layer and the adsorption material layer, the distance between the fluorescent dye of the biological sample and the feature array can be effectively reduced.

[0113] As another optional embodiment, as shown in FIG18 , a detection chip of the third structure mainly includes: a base material layer 11 , a feature array including a plurality of feature units 121 distributed on the base material layer 11 , and a feature array protection structure.

[0114] Among them, the feature array protection structure includes a feature array protection layer 132 and a sample adsorption layer formed by adsorption material. The feature array protection layer 132 is formed on the base material layer 11, the feature array is distributed in the feature array protection layer 132, and the sample adsorption layer is formed on the feature array protection layer 132.

[0115] The sample adsorption layer specifically includes a plurality of passivation regions 161 and a plurality of amination regions 162 . The amination regions 162 correspond one-to-one to the positions of the characteristic units along the extension direction of the characteristic units. The amination regions 162 are used to adsorb biological samples.

[0116] As an optional embodiment, the passivation region 161 can be formed by treating with a chemical reagent - hexamethyldisilazane (HDMS), and the amination region 162 is a region where amino groups are formed, which can connect DNA, etc. through amino groups. The specific process of treating with hexamethyldisilazane to form the passivation region 161 and the process of forming the amination region 162 can refer to the existing treatment process, so they will not be described here one by one.

[0117] In this embodiment, the feature array protective layer 132 can be further divided into a first feature array protective layer and a second feature array protective layer (not specifically distinguished in the figure), the feature array is distributed in the first feature array protective layer (whose thickness can be similar to the thickness of the feature unit 121), and the second feature array protective layer is formed on the first feature array protective layer and contacts the lower surface of the adsorption material layer 131.

[0118] The materials of the first characteristic array protection layer and the second characteristic array protection layer can be the same or different, and thus can be selected and adjusted accordingly according to actual needs or possible needs.

[0119] In this embodiment, the detection chip of the third structure is similar to the detection chip of the first structure in terms of the materials, sizes, distribution and shape of the feature arrays of each layer. Therefore, the specific parameters of the detection chip of the first structure can be referred to, and will not be described one by one.

[0120] In this embodiment, in the detection chip of the third structure, no sidewall material layer is required; the amino region formed on the feature array protective layer can be used to adsorb biological samples, thereby simplifying the overall structure of the detection chip and effectively simplifying the production process of the detection chip, improving production efficiency and saving production costs. The detection chip provided in this embodiment has the following major advantages:

[0121] 1. This embodiment can reduce system hardware costs: Using the detection chip of this embodiment, the local surface plasmon enhancement technology can greatly enhance the excitation power of the fluorescent dye on the DNA, thereby obtaining a higher fluorescence signal. In addition, because the local surface plasmon energy decays exponentially with distance, the laser background is low, which can effectively improve the signal-to-noise ratio of the imaging image. Therefore, for the same imaging quality, lower-power lasers, lower-cost components, and detectors with lower quantum efficiency and sensitivity can be selected, which can significantly reduce the overall hardware cost.

[0122] 2. This embodiment can increase sequencing throughput: For the same hardware system, since the detection chip of this embodiment can bring higher fluorescence signals and signal-to-noise ratios, while ensuring the same imaging quality, the exposure time can be reduced, thereby reducing the single run time of the sequencer and thereby improving the sequencing throughput.

[0123] 3. This embodiment can break through the sequencing throughput bottleneck of current traditional sequencing chips: In order to improve sequencing throughput, sequencers using traditional sequencing chips need to continuously reduce the single run time. Common methods include reducing exposure time and motion platform movement time, that is, using faster and better hardware equipment. However, the improvement of hardware equipment has its limits, so the improvement of sequencing throughput will encounter bottlenecks. The new detection chip of this embodiment can break through the sequencing throughput bottleneck of traditional sequencing chips.

[0124] As another embodiment, as shown in FIG19 , this embodiment provides a method for preparing the detection chip as in the above embodiment, which mainly includes the following steps:

[0125] Step 301: providing a base material layer;

[0126] Step 302: forming a feature array on the base material layer;

[0127] Step 303: forming a feature array protection structure on the base material layer so that the feature array is distributed within the feature array protection structure.

[0128] Specifically, as a specific embodiment, as shown in Figure 20, a layer of ultraviolet photoresist with a thickness of 300nm is spin-coated on an 8-inch silicon-based wafer, and then the photoresist is exposed and developed to obtain a feature array pattern. Next, a layer of metal (for example, Ag) is deposited, and the excess photoresist and metal are cleaned with acetone to obtain an Ag column layer as the feature array. Afterwards, a layer of SiO2 is electron-beam evaporated as a feature array protective layer to fill the gaps between the Ag columns and to be 10nm higher than the top of the Ag column. Then, a layer of TiO2 with a thickness of 10nm (as an adsorption material layer) and a layer of SiO2 with a thickness of 100nm (as a sidewall material layer) are evaporated. A layer of photoresist with a thickness of 300nm is spin-coated, the photoresist is exposed and developed, and then the SiO2 layer is dry-etched to obtain the final detection chip.

[0129] In this embodiment, the above-mentioned specific processing technology and process parameters are not specifically limited, and the processing technology and process parameters can be finely adjusted according to the actual processing equipment conditions.

[0130] The detection chip described above can be prepared using the method for preparing the detection chip provided in this embodiment, and has the beneficial effects described above.

[0131] Although the above describes specific embodiments of the present application, those skilled in the art will understand that these are merely illustrative and that the scope of protection of the present application is defined by the appended claims. Those skilled in the art may make various changes or modifications to these embodiments without departing from the principles and essence of the present application, and such changes and modifications shall fall within the scope of protection of the present application.

Claims

1. A detection chip, characterized in that: include: a base material layer; A feature array, comprising a plurality of feature units distributed on the base material layer; as well as A feature array protection structure is formed on the base material layer, and the feature array is distributed within the feature array protection structure, wherein the feature array protection structure at least includes an adsorption material for adsorbing biological samples. 2 . The detection chip according to claim 1 , wherein the plurality of characteristic units are regularly distributed on the base material layer. 3 . The detection chip according to claim 2 , wherein the plurality of characteristic units are regularly and periodically distributed on the base material layer.

4. The detection chip according to claim 1, further comprising a sidewall material layer formed on the feature array protection structure; The sidewall material layer includes a plurality of sidewall materials. The sample holes formed on the feature array protection structure by adjacent sidewall materials correspond one to one with the positions of the feature units along the extension direction of the feature units. The sample holes are used to accommodate the biological samples.

5. The detection chip according to claim 4, wherein the feature array protection structure comprises a feature array protection layer and an adsorption material layer formed by the adsorption material; The feature array protection layer is formed on the base material layer, the feature array is distributed in the feature array protection layer, the adsorption material layer is formed on the feature array protection layer, and the sidewall material layer is formed on the adsorption material layer.

6. The detection chip according to claim 5, wherein the feature array protection layer comprises a first feature array protection layer and a second feature array protection layer; The feature array is distributed in the first feature array protection layer, and the second feature array protection layer is formed on the first feature array protection layer and contacts the lower surface of the adsorption material layer.

7. The detection chip according to claim 5, wherein the thickness of the feature array protection layer is in the range of 5 nm to 200 nm; and / or The thickness of the adsorption material layer is in the range of 5 nm to 200 nm; and / or, The thickness of the sidewall material layer ranges from 10 nm to 1000 nm.

8. The detection chip according to claim 5, wherein the material of the feature array protection layer comprises any one or more of SiO2, Si3N4, and TiO2; and / or The material of the sidewall material layer includes any one or more of SiO2 and Si3N4.

9. The detection chip according to claim 4, wherein the feature array protection structure comprises an adsorption material layer formed by the adsorption material; The adsorption material layer is formed on the base material layer, the feature array is distributed in the adsorption material layer, and the sidewall material layer is formed on the adsorption material layer.

10. The detection chip according to claim 9, wherein the adsorption material layer comprises a first adsorption material layer and a second adsorption material layer; The feature array is distributed in the first adsorption material layer, and the second adsorption material layer is formed on the first adsorption material layer and contacts the lower surface of the sidewall material layer.

11. The detection chip according to claim 1, wherein the feature array protection structure comprises a feature array protection layer and a sample adsorption layer formed by the adsorption material; The feature array protection layer is formed on the base material layer, the feature array is distributed in the feature array protection layer, and the sample adsorption layer is formed on the feature array protection layer; in, The sample adsorption layer includes a plurality of passivation regions and a plurality of amination regions. The amination regions correspond one-to-one to the positions of the characteristic units along the extension direction of the characteristic units. The amination regions are used to adsorb the biological sample.

12. The detection chip according to claim 11, wherein the feature array protection layer comprises a first feature array protection layer and a second feature array protection layer; The feature array is distributed in the first feature array protection layer, and the second feature array protection layer is formed on the first feature array protection layer and contacts the lower surface of the sample adsorption layer. 13 . The detection chip according to claim 1 , wherein the base material layer is made of any one or more of silicon, quartz, or glass. 14 . The detection chip according to claim 1 , wherein the adsorption material comprises any one or more of TiN, TiO 2 , Ag, ZrO 2 , and ZnO. 15 . The detection chip according to claim 1 , wherein the feature array comprises a metal array, and the feature unit comprises a metal material. The detection chip according to claim 15 , wherein the metal material comprises any one or more of Ag, Au, Cu, and Al.

17. The detection chip according to any one of claims 1 to 14, wherein the plurality of characteristic units are periodically distributed on the base material layer in any one or more forms of polygonal, circular, and elliptical shapes; and / or The structure of the characteristic unit includes a cylinder or a sphere; and / or, The cross-sectional shape of the characteristic unit includes polygon, circle or ellipse.

18. The detection chip according to any one of claims 1 to 14, wherein the thickness of the characteristic unit is in the range of 20 nm to 300 nm; and / or The cross-sectional size of the characteristic unit ranges from 20 nm to 300 nm; and / or, The distance between the centers of two adjacent characteristic units ranges from 100 nm to 1000 nm.

19. The detection chip according to any one of claims 1 to 14, wherein a top surface of the feature array protection structure is higher than a top surface of any of the feature units.

20. A method for preparing a detection chip, characterized in that: The detection chip according to any one of claims 1 to 19 is prepared by the preparation method; The preparation method comprises: providing a base material layer; forming a feature array on the base material layer, wherein the feature array includes a plurality of feature units distributed on the base material layer; A feature array protection structure is formed on the base material layer so that the feature array is distributed within the feature array protection structure, wherein the feature array protection structure at least includes an adsorption material for adsorbing biological samples.

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