CN110591903A - Gene sequencing substrate, manufacturing method thereof and gene sequencing chip - Google Patents
Gene sequencing substrate, manufacturing method thereof and gene sequencing chip Download PDFInfo
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- CN110591903A CN110591903A CN201910889093.0A CN201910889093A CN110591903A CN 110591903 A CN110591903 A CN 110591903A CN 201910889093 A CN201910889093 A CN 201910889093A CN 110591903 A CN110591903 A CN 110591903A
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- AMGQUBHHOARCQH-UHFFFAOYSA-N indium;oxotin Chemical compound [In].[Sn]=O AMGQUBHHOARCQH-UHFFFAOYSA-N 0.000 claims description 7
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- YVTHLONGBIQYBO-UHFFFAOYSA-N zinc indium(3+) oxygen(2-) Chemical compound [O--].[Zn++].[In+3] YVTHLONGBIQYBO-UHFFFAOYSA-N 0.000 claims description 7
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Classifications
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- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
- C03C17/34—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions
- C03C17/36—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal
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- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
- C03C17/34—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions
- C03C17/36—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal
- C03C17/3602—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal the metal being present as a layer
- C03C17/3605—Coatings of the type glass/metal/inorganic compound
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- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
- C03C17/34—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions
- C03C17/36—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal
- C03C17/38—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal at least one coating being a coating of an organic material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
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- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C2218/00—Methods for coating glass
- C03C2218/30—Aspects of methods for coating glass not covered above
- C03C2218/32—After-treatment
- C03C2218/328—Partly or completely removing a coating
- C03C2218/33—Partly or completely removing a coating by etching
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- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C2218/00—Methods for coating glass
- C03C2218/30—Aspects of methods for coating glass not covered above
- C03C2218/34—Masking
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- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C2218/00—Methods for coating glass
- C03C2218/30—Aspects of methods for coating glass not covered above
- C03C2218/355—Temporary coating
Abstract
The embodiment of the application provides a gene sequencing substrate, a manufacturing method thereof and a gene sequencing chip. The gene sequencing substrate comprises: the device comprises a substrate, a metal reflecting layer positioned on one surface of the substrate, a functional layer positioned on one surface, far away from the substrate, of the metal reflecting layer, and a plurality of micropores penetrating through the functional layer, wherein the micropores are used as reaction tanks. The gene sequencing substrate provided by the embodiment can improve the fluorescence reflectivity of the gene detection chip by arranging the metal reflecting layer below the micropores which are used as the reaction tank, so that the sensitivity and the accuracy of gene sequencing are improved.
Description
Technical Field
The application relates to the technical field of gene sequencing, in particular to a gene sequencing substrate, a manufacturing method thereof and a gene sequencing chip.
Background
High-throughput Sequencing (High-throughput Sequencing) is capable of Sequencing hundreds of thousands to millions of DNA molecules at a time, and is therefore also referred to as Next Generation Sequencing (NGS).
The high-throughput sequencing technology is usually implemented by using a sequencing platform in combination with a gene sequencing chip to complete sequencing. In the existing gene detection chip, a nano well or a micro well is formed on a silicon chip as a reaction cell by using a semiconductor exposure and development process. However, the reflectivity of silicon is usually only 20% -40%, and the lower reflectivity causes the fluorescence emitted by the substance to be detected to have higher loss rate, thereby reducing the sensitivity and accuracy of gene sequencing.
Disclosure of Invention
The application provides a gene sequencing substrate, a manufacturing method thereof and a gene sequencing chip aiming at the defects of the existing mode, and aims to solve the technical problems of low sensitivity and low accuracy of gene sequencing caused by low fluorescence reflectivity of the gene sequencing chip based on a silicon wafer in the prior art.
In a first aspect, embodiments of the present application provide a gene sequencing substrate, comprising: a substrate; the metal reflecting layer is positioned on one surface of the substrate; the functional layer is positioned on one surface of the metal reflecting layer, which is far away from the substrate; and a plurality of micropores penetrating through the functional layer, wherein the micropores are used as reaction cells.
Optionally, the material of the functional layer comprises an imprint paste or an inorganic silicon material.
Optionally, the material of the metal reflective layer includes one or a combination of aluminum, silver, molybdenum and titanium.
Optionally, the substrate is a glass substrate or a quartz substrate.
Optionally, the gene sequencing substrate further comprises: and the transparent protective layer is positioned between the metal reflecting layer and the functional layer, and the material of the transparent protective layer comprises indium tin oxide and/or indium zinc oxide.
In a second aspect, the present application provides a gene sequencing chip, which includes the gene sequencing substrate and microbeads assembled in the micropores, wherein the microbeads carry biological probes.
In a third aspect, the present application provides a method for manufacturing a gene sequencing substrate, including:
forming a metal reflective layer on a substrate;
and forming a functional layer on the metal reflecting layer, and carrying out patterning treatment on the functional layer to form a plurality of micropores penetrating through the functional layer, wherein the micropores are used as reaction tanks.
Optionally, forming a functional layer on the metal reflective layer, and patterning the functional layer to form a plurality of micro holes penetrating through the functional layer, includes:
coating an imprinting adhesive, and curing the imprinting adhesive to serve as the functional layer;
and processing the imprinting glue by adopting a nano imprinting process to form the micropores penetrating through the functional layer.
Optionally, patterning the functional layer to form a plurality of micro-holes penetrating through the functional layer, including:
depositing an inorganic silicon material layer on the metal reflecting layer to serve as the functional layer;
forming an imprinting glue layer on the functional layer;
forming a plurality of through holes penetrating through the imprinting glue by adopting a nano-imprinting process, or forming a plurality of grooves on the imprinting glue by adopting the nano-imprinting process;
etching the inorganic silicon material layer by using the imprinting glue for forming a plurality of through holes or a plurality of grooves as a mask so as to form a plurality of micropores penetrating through the inorganic silicon material layer;
and removing the stamping glue.
Optionally, the manufacturing method further includes: before the functional layer is formed, a transparent protective layer is formed on the metal reflecting layer, and the material of the transparent protective layer comprises indium tin oxide and/or indium zinc oxide.
The technical scheme provided by the embodiment of the application has the following beneficial technical effects:
according to the gene sequencing substrate, the manufacturing method thereof and the gene sequencing chip provided by the embodiment, the metal reflecting layer is arranged below the micropores serving as the reaction tanks, so that the fluorescence reflectivity of the gene detection chip can be improved, and the sensitivity and the accuracy of gene sequencing are improved.
Additional aspects and advantages of the present application will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the present application.
Drawings
The foregoing and/or additional aspects and advantages of the present application will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a schematic diagram showing a structure of a gene sequencing chip according to the related art;
FIG. 2 is a schematic diagram of a gene sequencing substrate according to an embodiment of the present disclosure;
FIG. 3 is a schematic diagram showing the structure of another gene sequencing substrate according to the present embodiment;
FIG. 4 is a schematic diagram of a gene sequencing chip according to an embodiment of the present disclosure;
FIG. 5 is a schematic flow chart of a method for manufacturing a gene sequencing substrate according to an embodiment of the present disclosure;
fig. 6 is a schematic process diagram of step S1 in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
FIG. 7 is a schematic process diagram of step S2 in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
FIG. 8 is a schematic flow chart of step S2 in the method for manufacturing a gene sequencing substrate according to the present application;
fig. 9 is a schematic process diagram of step S202 in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
FIG. 10 is a schematic flow chart illustrating a step S2 of the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
fig. 11 is a process diagram of step S201' in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
fig. 12 is a process diagram of step S202' in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
fig. 13 is a schematic process diagram of step S203' in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
fig. 14 is a schematic process diagram of step S204' in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application;
fig. 15 is a schematic process diagram of step S205' in the method for manufacturing a gene sequencing substrate according to the embodiment of the present application.
Reference numerals:
1' -a silicon wafer; 1' 0-groove;
2-a microbead;
1-a substrate;
2-a metal reflective layer;
3-a functional layer; 31-microwell;
3 a-impression glue; 30 a-microwell;
3 b-a layer of inorganic silicon material; 30 b-microwells;
4-a transparent protective layer;
5-impression glue;
m-imprint template.
Detailed Description
Reference will now be made in detail to the present application, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar parts or parts having the same or similar functions throughout. In addition, if a detailed description of the known art is not necessary for illustrating the features of the present application, it is omitted. The embodiments described below with reference to the drawings are exemplary only for the purpose of explaining the present application and are not to be construed as limiting the present application.
It will be understood by those within the art that, unless otherwise defined, all terms (including 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 belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the prior art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.
As used herein, the singular forms "a", "an", "the" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and/or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. It will be understood that when an element is referred to as being "connected" or "coupled" to another element, it can be directly connected or coupled to the other element or intervening elements may also be present. Further, "connected" or "coupled" as used herein may include wirelessly connected or wirelessly coupled. As used herein, the term "and/or" includes all or any element and all combinations of one or more of the associated listed items.
The inventor of the present application considers that, in the existing gene detection chip, a nano well or a micro well is formed on a silicon wafer as a reaction cell by using a semiconductor exposure and development process, and the sensitivity and the accuracy of gene sequencing are reduced due to the low light reflectivity of the silicon wafer.
Specifically, as shown in fig. 1, in the gene detection chip in the related art, a silicon wafer 1 'is used as a substrate material, a well structure 10' serving as a reaction tank is formed on one side of the silicon wafer 1 ', and a bead carrying a biological probe is assembled in the well structure 10', so that when the biological probe is combined with a gene to be detected having different fluorescence characteristics, the biological probe can emit fluorescence of different colors, and the gene sequence can be read by detecting the fluorescence. However, the reflectivity of the silicon material is usually only 20% -40%, and the lower reflectivity enables the fluorescence emitted by the substance to be detected to have higher loss rate, thereby reducing the sensitivity and accuracy of gene sequencing.
The application provides a gene sequencing substrate, a manufacturing method thereof and a gene sequencing chip, and aims to solve the technical problems in the prior art.
The following describes the technical solutions of the present application and how to solve the above technical problems with specific embodiments.
This example provides a gene sequencing substrate, as shown in fig. 2, comprising:
a substrate 1;
a metal reflective layer 2 on one side of the substrate 1;
the functional layer 3 is positioned on one surface of the metal reflecting layer 2, which is far away from the substrate 1; and
a plurality of micropores 30 as reaction cells penetrating the functional layer 3.
The gene sequencing substrate provided by the embodiment can improve the fluorescence reflectivity of a gene detection chip by arranging the metal reflecting layer 2 below the micropore 30 which is used as a reaction tank, so that the sensitivity and the accuracy of gene sequencing are improved.
Specifically, the substrate 1 is a glass substrate, a quartz substrate, or the like, and has a certain rigidity, so that a good supporting function can be achieved. And the glass substrate, the quartz substrate and other substrates with certain rigidity have higher flatness, and can ensure that when the microbeads are filled into the micropores 30 (i.e. the beads), the microbeads are tiled into a single layer without agglomeration. On the basis, the fluorescence signal intensity spectrum can truly reflect the test result in the subsequent laser detection. In addition, since a glass substrate, a quartz substrate, or the like can be processed into a substrate having an area much larger than that of a wafer (silicon material), an existing production line of a Liquid Crystal Display (LCD) can be used, so that not only can the investment cost of production equipment be saved, but also mass production can be realized.
Alternatively, as shown in fig. 2, the material of the functional layer 3 is an imprint paste or an inorganic silicon material. Specifically, the imprinting adhesive may be a thermosetting imprinting adhesive or an ultraviolet curing imprinting adhesive, the imprinting adhesive is used as a material of the functional layer 3, the nano-imprinting process may be used to form the micro-pores 30, the production process is simple, the production cost is low, and the large-scale production can be realized. The inorganic silicon material can be one or a combination of silicon, silicon oxide, silicon nitride and silicon oxynitride, and the inorganic silicon material is used as the material of the functional layer 3, so that the biological reagent has good tolerance and wider applicability.
Optionally, as shown in fig. 2, the material of the metal reflective layer 2 includes one or a combination of aluminum, silver, molybdenum, and titanium. Taking the metal reflecting layer 2 as an aluminum film as an example, the full-spectrum reflectivity of the aluminum film is 75% -90%, which is obviously superior to 20% -40% of that of a silicon wafer, and the fluorescence intensity to be detected can be improved, so that the sensitivity and accuracy of gene sequencing are improved.
Alternatively, the size of the micro-pores 30 is nano-scale or micro-scale, and the existing process can control the micro-pores to be several nanometers to several hundred micrometers, and typically, the size of the micro-pores 30 is controlled to be between 1nm and 100 μm. Specifically, the size of the microwell 30 is designed according to the size of the microbeads required to fit in the microwell 30, for example, the size of a certain kind of microbeads carrying biological probes, which are provided by illumina, inc, is 1 micrometer, and thus the size of the microwell 30 on the gene sequencing substrate suitable for such microbeads should be set to 1 micrometer.
Referring to fig. 3, in addition, the gene sequencing substrate provided in this embodiment further includes: and the transparent protective layer 4 is positioned between the metal reflecting layer 3 and the functional layer 3, and the material of the transparent protective layer 4 comprises one or more of indium tin oxide and indium zinc oxide. The transparent protective layer 4 protects the metal reflective layer 2 from corrosion of the metal reflective layer 3 by a reagent injected into the micro holes 30, or water or the like introduced into the micro holes 30. Meanwhile, the good tolerance of the micropores to biological reagents can be further improved.
Specifically, whether to set the transparent protection layer 4 may be selected according to the material of the metal reflection layer 2 and the object to be detected to which the gene detection chip is applied, for example, the material of the metal reflection layer 2 is aluminum, and the reagent of the object to be detected contains an alkaline substance, and the transparent protection layer 4 needs to be set in order to avoid the reaction between the aluminum and the alkaline substance. The transparent protective layer 4 may not be provided when the metal reflective layer 2 does not react with the object to be detected or the reagent of the object to be detected.
Based on the same light-emitting concept, this embodiment provides a gene sequencing chip, please refer to fig. 4, the gene detection chip includes the gene sequencing substrate in the above embodiment and a bead assembled in the micro-well 30, the bead carries a biological probe. Since the gene sequencing chip provided in this embodiment includes the gene sequencing substrate in the above embodiments, the beneficial effects of the gene sequencing substrate can be achieved, and details are not repeated herein.
Based on the same light-emitting concept, this embodiment provides a method for manufacturing a gene sequencing substrate, as shown in fig. 5, the method includes:
s1: a metal reflective layer is formed on a substrate. As shown in fig. 6, specifically, one or a combination of materials such as aluminum, aluminum alloy, silver, molybdenum, titanium, and the like is deposited on the substrate 1 to form the metal reflective layer 2. The film formed by the metals has higher light reflection characteristic, and taking an aluminum film as an example, the full-spectrum reflectivity of the aluminum film is 75-90%, which is obviously better than that of a silicon wafer by 20-40%.
S2: referring to fig. 7 and 2, a functional layer 3 is formed on the metal reflective layer 2, and the functional layer 3 is patterned to form a plurality of micro holes 30 penetrating through the functional layer, wherein the micro holes 30 serve as reaction cells.
The gene sequencing substrate provided by the embodiment can improve the fluorescence reflectivity of a gene detection chip by arranging the metal reflecting layer 2 below the micropore 30 which is used as a reaction tank, so that the sensitivity and the accuracy of gene sequencing are improved.
Specifically, the substrate 1 is a glass substrate, a quartz substrate, or the like, and has a certain rigidity, so that a good supporting function can be achieved. And the glass substrate, the quartz substrate and other substrates with certain rigidity have higher flatness, and can ensure that when the microbeads are filled into the micropores 30 (i.e. the beads), the microbeads are tiled into a single layer without agglomeration. On the basis, the fluorescence signal intensity spectrum can truly reflect the test result in the subsequent laser detection. In addition, since a glass substrate, a quartz substrate, or the like can be processed into a substrate having an area much larger than that of a wafer (silicon material), an existing production line of a Liquid Crystal Display (LCD) can be used, so that not only can the investment cost of production equipment be saved, but also mass production can be realized.
Optionally, referring to fig. 8 and 9, in the method for manufacturing a gene sequencing chip provided in this embodiment, step S2 includes:
s201: an imprint paste 3a is coated on the metal reflective layer 2, and the imprint paste 3a is cured to serve as a functional layer. Specifically, the curing of the imprinting paste 3a may be performed by a heat curing process or an ultraviolet curing process, depending on the type of the imprinting paste 3 a.
S202: the imprinting paste 3a is patterned using a nano-imprinting process to form micro-holes 30a penetrating the imprinting paste 3 a. Specifically, in the nano-imprint process, a prefabricated suppression template M is used to imprint on the imprint glue 3a, and then the imprint template M is separated from the imprint glue 3a, so that the micro-holes 30a can be formed.
According to the manufacturing method of the gene sequencing substrate, the imprinting glue 3a is used as the material of the functional layer, the micropores 30a can be formed on the imprinting glue 3a through the nano imprinting technology, the manufacturing method is simple in process, the cost can be reduced at present, and the manufacturing method is suitable for large-scale production.
Optionally, referring to fig. 10, in the method for manufacturing a gene sequencing substrate provided in this embodiment, step S2 includes:
s201': as shown in fig. 11, an inorganic silicon material layer 3b is deposited as a functional layer on the metal reflective layer 2. Specifically, one or a combination of materials such as silicon, silicon oxide, silicon nitride, and silicon oxynitride may be deposited on the metal reflective layer 2 to form the inorganic silicon material layer 3 b.
S202': as shown in fig. 12, the imprinting paste 5 is coated on the inorganic silicon material layer 3b, and the imprinting paste 5 is cured. Specifically, the curing of the imprint glue 5 may be performed by a heat curing process or an ultraviolet curing process, depending on the type of the imprint glue 5.
S203': as shown in fig. 13, a plurality of grooves 50 are formed on the imprinting glue using a nano-imprinting process. Of course, a nano-imprint process may be used to form a plurality of through holes through the imprint resist 5.
S204': as shown in fig. 14, the inorganic silicon material layer 3b is dry-etched using the imprint paste 5 forming the plurality of grooves 50 or the plurality of through holes as a mask to form a plurality of micro holes 30b penetrating the inorganic silicon material layer 3 b. Specifically, the process parameters of the dry etching may be selected according to the material of the inorganic silicon material layer 3b and the thickness of the inorganic silicon material layer 3 b.
S205': as shown in fig. 15, the imprint glue 5 is removed.
According to the manufacturing method of the gene sequencing substrate, the functional layer for forming the micropores is manufactured by adopting the inorganic silicon material, so that the formed micropores 30b have good tolerance to biological reagents and can be suitable for more extensive objects to be detected, and the imprinting glue treated by the nano imprinting process is used as a mask, so that the process is simple, the cost can be reduced, and the method is suitable for large-scale production.
Optionally, referring to fig. 3, the method for manufacturing a gene sequencing substrate provided in this embodiment further includes: before the functional layer 3 is formed, a transparent protective layer 4 is formed on the metal reflective layer 2, and a material of the transparent protective layer 4 includes indium tin oxide and/or indium zinc oxide. Specifically, indium tin oxide and/or indium zinc oxide material may be deposited on the metal reflective layer 2 to form the transparent protective layer 4. The transparent protective layer 4 protects the metal reflective layer 2 to prevent corrosion of the metal reflective layer by a reagent injected into the micro-holes, or by water or the like entering the micro-holes. Meanwhile, the good tolerance of the micropores to biological reagents can be further improved.
By applying the embodiment of the application, at least the following beneficial effects can be realized:
according to the gene sequencing substrate, the manufacturing method thereof and the gene sequencing chip provided by the embodiment, the metal reflecting layer is arranged below the micropores serving as the reaction tanks, so that the fluorescence reflectivity of the gene detection chip can be improved, and the sensitivity and the accuracy of gene sequencing are improved.
Those of skill in the art will appreciate that the various operations, methods, steps in the processes, acts, or solutions discussed in this application can be interchanged, modified, combined, or eliminated. Further, other steps, measures, or schemes in various operations, methods, or flows that have been discussed in this application can be alternated, altered, rearranged, broken down, combined, or deleted. Further, steps, measures, schemes in the prior art having various operations, methods, procedures disclosed in the present application may also be alternated, modified, rearranged, decomposed, combined, or deleted.
In the description of the present application, it is to be understood that the terms "center", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, should not be construed as limiting the present invention.
The terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified.
In the description of the present application, it is to be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
In the description herein, particular features, structures, materials, or characteristics may be combined in any suitable manner in any one or more embodiments or examples.
It should be understood that, although the steps in the flowcharts of the figures are shown in order as indicated by the arrows, the steps are not necessarily performed in order as indicated by the arrows. The steps are not performed in the exact order shown and may be performed in other orders unless explicitly stated herein. Moreover, at least a portion of the steps in the flow chart of the figure may include multiple sub-steps or multiple stages, which are not necessarily performed at the same time, but may be performed at different times, which are not necessarily performed in sequence, but may be performed alternately or alternately with other steps or at least a portion of the sub-steps or stages of other steps.
The foregoing is only a partial embodiment of the present application, and it should be noted that, for those skilled in the art, several modifications and decorations can be made without departing from the principle of the present application, and these modifications and decorations should also be regarded as the protection scope of the present application.
Claims (10)
1. A gene sequencing substrate, comprising:
a substrate;
the metal reflecting layer is positioned on one surface of the substrate;
the functional layer is positioned on one surface of the metal reflecting layer, which is far away from the substrate; and
and the micropores penetrate through the functional layer and are used as reaction cells.
2. The gene sequencing substrate of claim 1, wherein the material of the functional layer comprises an imprinted gel or an inorganic silicon material.
3. The gene sequencing substrate of claim 1, wherein the material of the metal reflective layer comprises one or a combination of aluminum, silver, molybdenum and titanium.
4. The gene sequencing substrate of claim 1, wherein the substrate is a glass substrate or a quartz substrate.
5. The gene sequencing substrate of any one of claims 1-4, further comprising:
and the transparent protective layer is positioned between the metal reflecting layer and the functional layer, and the material of the transparent protective layer comprises indium tin oxide and/or indium zinc oxide.
6. A gene sequencing chip comprising the gene sequencing substrate according to any one of claims 1 to 5 and microbeads assembled in the microwells, the microbeads carrying biological probes.
7. A method for manufacturing a gene sequencing substrate, comprising:
forming a metal reflective layer on a substrate;
and forming a functional layer on the metal reflecting layer, and carrying out patterning treatment on the functional layer to form a plurality of micropores penetrating through the functional layer, wherein the micropores are used as reaction tanks.
8. The method of claim 7, wherein forming a functional layer on the metallic reflective layer and patterning the functional layer to form a plurality of micro-holes through the functional layer comprises:
coating an imprinting adhesive, and curing the imprinting adhesive to serve as the functional layer;
and processing the imprinting glue by adopting a nano imprinting process to form the micropores penetrating through the functional layer.
9. The method of claim 7, wherein patterning the functional layer to form a plurality of pores through the functional layer comprises:
depositing an inorganic silicon material layer on the metal reflecting layer to serve as the functional layer;
forming an imprinting glue layer on the functional layer;
forming a plurality of through holes penetrating through the imprinting glue by adopting a nano-imprinting process, or forming a plurality of grooves on the imprinting glue by adopting the nano-imprinting process;
etching the inorganic silicon material layer by using the imprinting glue for forming a plurality of through holes or a plurality of grooves as a mask so as to form a plurality of micropores penetrating through the inorganic silicon material layer;
and removing the stamping glue.
10. The method of manufacturing according to any one of claims 7 to 9, further comprising:
before the functional layer is formed, a transparent protective layer is formed on the metal reflecting layer, and the material of the transparent protective layer comprises indium tin oxide and/or indium zinc oxide.
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Application publication date: 20191220 |