Manufacturing method of biochip, product of biochip and detection method using biochip
Patent Information
- Application Number
- CN202380080264.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-01-16
- Publication Date
- 2025-06-27
AI Technical Summary
Existing biochips have high density, high manufacturing difficulty and high manufacturing costs, and require special instruments for storage, use and interpretation. They are difficult to popularize and cost, and lack low-cost and efficient manufacturing methods.
By generating multiple observation holes on the substrate, and setting fixings around these holes, the reaction sheet is fixed on the bottom surface of the substrate, and the deformation of the fixings (such as melting by heating) is used to further fix the reaction sheet, using the reaction sheet that has been cut in advance and Fasteners enable the combination of different reaction components, reducing manufacturing costs and improving flexibility of use.
A low-cost, simple-to-manufacture biochip is realized, which can arrange different types of reaction sheets on a single substrate, saving manufacturing costs, and improving the flexibility of use of the biochip and the accuracy and speed of analysis results.
Smart Images
Figure CN120225876A_ABST
Abstract
Description
Method for manufacturing biochip and its product, and detection method using the same Technical Field
[0001] The present disclosure relates to a method for manufacturing a biochip, specifically, a method for manufacturing a biochip in which a plurality of reaction chips are disposed on a substrate and fixed with fixing members on the substrate. Background Art
[0002] Biochips originated in the 1980s. While various electronic industries were miniaturizing their products, related industries such as life sciences and bioinformatics also realized the progress that could be brought about by miniaturizing biochemical analysis.
[0003] Broadly speaking, a biochip is a device that utilizes principles from molecular biology, analytical chemistry, and biochemical reactions to miniaturize reaction components used to analyze samples and affix them to a substrate made of glass, silicon, or plastic. The objects analyzed can be genes, proteins, cell tissues, and more. By miniaturizing reaction components, the accuracy, speed, and integrity of analytical results can be improved, while requiring significantly less sample and reagent volume than traditional analytical methods.
[0004] Specifically, biochips can be divided into two categories: microarray chips that plant high-density biological probes on a small area of the chip, and microprocessor chips that process biological samples and perform reaction analysis on the chip. Among them, the production methods of microarray chips are divided into: (1) in situ synthesis method, that is, direct synthesis on a carrier; and (2) placement method, that is, placing pre-synthesized probes on a carrier. The in situ synthesis method is suitable for the production of high-density DNA chips, but it is expensive and has limited efficiency, and its application range is relatively limited. In comparison, the placement method is relatively low in cost and has a wider range of applications, making it suitable for use in academic institutions or general enterprises.
[0005] However, currently used biochips have high density, difficulty in manufacturing, and high costs, making them difficult to widely use. Furthermore, their storage, use, and interpretation require specialized instruments. Therefore, a biochip and its manufacturing method are needed that can improve the accuracy, speed, and integrity of analysis results, while requiring less sample and reagent volume than traditional analysis methods and at a lower cost.
[0006] Summary of the Invention
[0007] To achieve the above-mentioned objectives, the present application provides a method for manufacturing a biochip, comprising: manufacturing a substrate; generating a plurality of observation holes penetrating the substrate on the substrate; setting a plurality of fixing members on the bottom surface of the substrate around each of the plurality of observation holes; setting a plurality of reaction plates on one side of the bottom surface respectively corresponding to the plurality of observation holes; and fixing the plurality of reaction plates by the plurality of fixing members.
[0008] Preferably, the method further comprises deforming the plurality of fixing members to fix the plurality of reaction sheets.
[0009] Preferably, the deformation is performed by heating and melting the plurality of fixing members.
[0010] Preferably, it further comprises arranging a reaction component on a reaction master sheet, and cutting the reaction master sheet into a plurality of reaction sheets.
[0011] Preferably, each of the reaction pieces includes two or more reaction components.
[0012] Preferably, the minimum distance between the centers of the reaction pieces is 1 to 10 mm.
[0013] Preferably, the fixing member is in the shape of an L-shaped column, a cylinder, a polygonal column, a cone, a cylinder, a sphere, a polyhedron, a ring or a combination thereof.
[0014] In order to achieve another object of this application, a biochip is provided, which is manufactured using the aforementioned biochip manufacturing method.
[0015] Preferably, each of the observation holes is surrounded by at least one corresponding fixing member.
[0016] Preferably, the substrate is made of silicon, glass, polymer material or ceramic.
[0017] Preferably, the fixing member is made of plastic, silicone or rubber.
[0018] To achieve another object of this application, a method for detecting samples using the aforementioned biochip is provided, comprising: adding samples to the reaction plate through observation holes; and observing the results through the observation holes.
[0019] Through the aforementioned technical features, the biochip manufacturing method provided herein enables the production of biochips with low cost and simple manufacturing methods. Furthermore, by pre-cutting reaction chips and fixtures, different types of reaction chips can be arranged on a single substrate, allowing for different reaction component combinations based on different needs, thereby further reducing manufacturing costs and increasing the flexibility of biochip usage. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] In the following detailed description, in order to explain the present application, many specific details are provided so that the disclosed embodiments can be thoroughly understood. However, it is apparent that one or more embodiments can be implemented without these specific details. In other cases, in order to simplify the drawings, conventional structures and processes are shown in a schematic manner.
[0021] FIG1 is a flow chart of a biochip manufacturing method according to an embodiment of the present application.
[0022] FIG. 2 is a schematic diagram of a substrate used in the biochip manufacturing method according to the first embodiment of the present application.
[0023] FIG3 is a schematic diagram of forming an observation hole on a substrate in the biochip manufacturing method according to the first embodiment of the present application.
[0024] FIG4 is an oblique view of a biochip including a conical fixing member according to the first embodiment of the present application.
[0025] FIG. 5 is a schematic side view of a cross section of the substrate along the sectional line AA′ of FIG. 4 .
[0026] FIG6 is a schematic side view of a cross section of a reaction chip disposed on a substrate in the biochip manufacturing method according to the first embodiment of the present application.
[0027] FIG. 7 is a schematic diagram illustrating melting a fixing member to further fix the reaction chip in the biochip manufacturing method according to the first embodiment of the present application.
[0028] FIG8 is a perspective view of a biochip including a mushroom-shaped fixing member according to a second embodiment of the present application.
[0029] FIG. 9 is a schematic side view of a cross section of the substrate along the BB′ line according to FIG. 8 .
[0030] FIG. 10 is a schematic side view of a cross section of a reaction chip disposed on a substrate in a biochip manufacturing method according to a second embodiment of the present application. DETAILED DESCRIPTION
[0031] The following is a detailed description of the embodiments with reference to the relevant drawings. However, the embodiments can be implemented in different forms, but this is not the only form of implementing or using the specific embodiments of the application requested by this application, and should not be understood as a limitation on the above embodiments. The features of multiple specific embodiments and the method steps and their sequence for constructing and operating these specific embodiments are covered in the implementation method. However, other specific embodiments can also be used to achieve the same or equal functions and step sequences. On the contrary, the embodiments are provided so that this specification can be thoroughly and completely disclosed to fully express the spirit of the application requested by this application to those skilled in the art to which this application belongs. Similar component symbols in the drawings refer to similar components. In the following description, conventional functions or structures will not be described in detail so as not to repeat unnecessary details in the embodiments.
[0032] Unless otherwise defined, all technical terms and terminology used herein have the same meaning as commonly understood by those skilled in the art to which this application belongs. In the event of a conflict, the present specification, including definitions, will control.
[0033] Unless otherwise specified, singular nouns used in this specification include plural nouns, and plural nouns include singular nouns. Furthermore, in this specification and claims, expressions such as "at least one" and "one or more" have the same meaning, meaning one, two, three, or more.
[0034] Although the numerical ranges and parameters used to define the broader scope of the claimed invention are approximate, the numerical values presented herein have been presented as precisely as possible in the specific examples. However, any numerical value inherently and inevitably contains standard deviations resulting from individual testing methods. As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specified value or range. Alternatively, the term "about" means that the actual value falls within an acceptable standard error of the mean, as determined by one skilled in the art. Except in the examples, or unless expressly indicated otherwise, all ranges, amounts, values, and percentages used herein (e.g., to describe material amounts, time periods, temperatures, operating conditions, quantitative ratios, and the like) are to be understood as modified by the word "about." Therefore, unless otherwise indicated, the numerical parameters disclosed in this specification and claims are approximate and may be modified as needed. At a minimum, these numerical parameters should be understood to include the number of significant digits indicated and to apply normal rounding. Herein, numerical ranges are expressed from one end point to another or between two end points; unless otherwise stated, the numerical ranges described herein include the end points.
[0035] First, please refer to FIG1 , which is a flow chart of a biochip manufacturing method according to an embodiment of the present application.
[0036] According to one embodiment of the present application, a method for manufacturing a biochip is provided, comprising:
[0037] S1: step of manufacturing substrate,
[0038] S2: a step of generating a plurality of observation holes penetrating the substrate on the substrate,
[0039] S3: a step of arranging a plurality of fixing members on the bottom surface of the substrate around each of the plurality of observation holes,
[0040] S4: a step of arranging a plurality of reaction plates on one side of the bottom surface corresponding to the plurality of observation holes, and
[0041] S5: a step of fixing the plurality of reaction pieces by using the plurality of fixing members.
[0042] According to another embodiment of the present application, after the plurality of reaction sheets are arranged on the bottom surface of the substrate, the step of melting the fixing parts to fix the reaction sheets more firmly on the bottom surface of the substrate is further included. The method of melting the fixing parts includes but is not limited to heating. Any method that can deform the fixing parts and prevent the reaction sheets from falling off can be used. Specifically, for example, the metal sheet can be heated and then pressed flat against the fixing parts to flatten them. According to another embodiment of the present application, after the reaction sheets are arranged on the bottom surface of the substrate, they can be further fixed with resin or glue to increase the firmness or sealing of the reaction sheets.
[0043] According to one embodiment of the application, the manufacture method of biochip can further include that reaction assembly (such as biological probe) is arranged on reaction master, and is cut into the step of multiple reaction slices.The material of reaction master can be paper or plastic, preferably plastic, and it has elasticity and is easy to arrange.The method that reaction assembly is set in reaction master can comprise photomask type, pinpoint type, inkjet type and piezoelectric type, and can adopt suitable mode to arrange reaction assembly according to demand difference.The mode of cutting reaction master can comprise model tool cutting or laser cutting etc., can be adjusted according to the different biological probe types required.
[0044] According to one embodiment of the present application, the reaction sheet can be circular, square, rectangular, or hexagonal, and can be adjusted as needed, preferably circular. According to one embodiment of the present application, the reaction sheet can be circular with a diameter of 1 to 3 mm, preferably circular with a diameter of 1.5 mm. According to another embodiment of the present application, the reaction sheet can be square with a side length of 1 to 3 mm, preferably square with a side length of 1.5 mm.
[0045] According to one embodiment of the present application, the fixing member may be in the shape of an L-shaped column, a cylinder, a polygonal column, a cone, a cylinder, a sphere, a polyhedron, a ring, or a combination thereof. Preferably, the fixing member may be a cone. According to another embodiment of the present application, the fixing member may be in the shape of a cylinder or a polygonal column where it contacts the bottom surface of the substrate, and may be in the shape of a sphere or a polyhedron where it does not contact the substrate.
[0046] According to one embodiment of the present application, the reaction component can be a protein probe, such as an antigen, an antibody, or a nucleic acid probe, such as a DNA probe or an RNA probe, but is not limited thereto. A person skilled in the art of the present application can set the required or desired reaction components according to the common knowledge in the art. By setting different reaction components (reaction chips) on a single biochip, the testing of the same or multiple samples at different stages can be integrated on the same biochip, making it easier for users to order the reactions they want to perform on a biochip and observe the results.
[0047] The biochip can be manufactured by the above-mentioned biochip manufacturing method. According to one embodiment of the present application, the observation hole on the biochip can be surrounded by at least one fixing member, preferably, can be surrounded by four fixing members. On the other hand, the substrate of the biochip can be made of silicon, glass, polymer material or ceramic, preferably made of polymer material, such as plastic. According to one embodiment of the present application, the fixing member can be made of plastic, silicone or rubber. Specifically, any material that is easy to shape and can be set on the substrate can be suitable for the fixing member, preferably plastic. According to another embodiment of the present application, after the reaction piece is set on the fixing member, the fixing member is deformed to further fix the reaction piece, for example, the fixing member is heated and melted to cause it to deform, or the fixing member is deformed by extrusion.
[0048] According to one embodiment of the present application, a method for detecting a sample using a biochip manufactured using the aforementioned method is provided, comprising dripping the sample into an observation hole from the opposite side of the bottom surface where the fixing member is provided, so that the sample contacts the reaction plate for reaction, and observing the result through the observation hole by the naked eye or an instrument.
[0049] The biochip manufacturing method, biochip, and sample detection method provided in this application are described in detail below based on specific embodiments and drawings.
[0050] Please refer to Figures 2 to 6. Figure 2 is a schematic diagram of a substrate used in the biochip manufacturing method according to the first embodiment of the present application. Figure 3 is a schematic diagram of generating an observation hole on a substrate in the biochip manufacturing method according to the first embodiment of the present application. Figure 4 is an oblique view of a biochip including a conical fixing member according to the first embodiment of the present application. Figure 5 is a side view schematic diagram of a cross section along the tangent line A-A' of the substrate according to Figure 4. Figure 6 is a side view schematic diagram of a cross section of a reaction plate arranged on a substrate in the biochip manufacturing method according to the first embodiment of the present application. Figure 7 is a schematic diagram of melting a fixing member to further fix the reaction plate in the biochip manufacturing method according to the first embodiment of the present application.
[0051] According to a first embodiment of the present application, a substrate 1 as shown in Figure 2 is prepared. In this embodiment, substrate 1 is formed of plastic. Next, as shown in Figure 3, a plurality of observation holes 2 are formed through substrate 1. In this embodiment, the diameter of the observation holes 2 is 1.1 mm, and the distance between adjacent observation holes is 1.54 mm.
[0052] Please refer to Figure 4. The bottom of Figure 4 is the front of the substrate 1, and the top is the bottom of the substrate 1. In the first embodiment, a plurality of conical fixing parts 3 are formed on the bottom side of the substrate 1, and the fixing parts 3 are made of plastic. In addition, since the sample will be added from the front of the biochip when dripping, the cross-section of the observation hole 2 is set to be narrow at the top and wide at the bottom as shown in Figure 4 to facilitate sample addition. On the other hand, this setting can prevent the reaction piece 4 from falling into the observation hole 2 when the reaction piece 4 is laid out. In this embodiment, the inner diameter (bottom side) of the observation hole 2 is 1.1 mm, and the outer diameter (front side) is 1.2 mm. The center distance between the observation holes 2 in the same row or column is 1.54 mm, and the center distance between the conical fixing parts 3 in the same row or column is 1.54 mm. The substrate 1 with the conical fixing parts 3 is shown in Figure 5.
[0053] Next, referring to Figure 6 , a reaction sheet 4 is placed on the bottom surface of substrate 1. In this embodiment, reaction sheet 4 is a circular plastic sheet with a diameter of 1.4 mm. The reaction component mounted on top is a protein probe. After placement of reaction sheet 4, the tips of the conical plastic fixtures 3 are melted and flattened with an iron to create a fixing portion 5, as shown in Figure 7 . This secures reaction sheet 4 between the four conical fixtures 3 and between the fixing portion 5 and substrate 1.
[0054] The second embodiment of the present application is described below with reference to Figures 8 to 10. Figure 8 is an oblique view of a biochip including a mushroom-shaped fixture according to the second embodiment of the present application. Figure 9 is a schematic side view of a cross section of the substrate along the BB' line according to Figure 8. Figure 10 is a schematic side view of a cross section of a reaction chip disposed on a substrate in the biochip manufacturing method according to the second embodiment of the present application.
[0055] The biochip manufacturing method of the second embodiment of this application shares similarities with the first embodiment and will not be described in detail. The difference lies in the mushroom-shaped fixture 3a, as shown in Figures 8 and 9. The mushroom-shaped fixture 3a facilitates insertion and fixation of the reaction chip 4. In the second embodiment, the reaction chip 4 is made of a slightly elastic plastic sheet. The biochip after the reaction chip 4 is installed is shown in Figure 10.
[0056] In the third embodiment of the present application, the structures of the substrate, observation aperture, and fixing member are similar to those described in the first and second embodiments. However, in the third embodiment, the bioprobes are applied to a paper reaction chip using an inkjet method. The reaction chip is then cut into circular pieces with a diameter of 1.4 mm by laser cutting, resulting in a reaction chip 4. Next, a placement machine is used to absorb the back of the reaction chip 4, where the bioprobes are not applied, and the reaction chip is fixed between fixing members 3 and 3a. This allows different types of reaction chips 4 to be placed on the same biochip according to experimental requirements, thereby achieving cost savings. According to one embodiment of the present application, after the fixing members 3 are flattened to form the fixing portion 5, they are then tightly fitted together, sealing the reaction chip 4 between the fixing portion 5 and the substrate 1. In this case, when observing the results through the observation aperture 2, an external light source can be used to assist in observation. According to another embodiment of the present application, after the fixing members 3 are flattened to form the fixing portion 5, they are not tightly fitted together. When observing the results through the observation aperture 2, a light source can be applied from the bottom surface of the substrate 1 to assist in observation.
[0057] According to a fourth embodiment of the present application, a method for detecting a sample using the biochip prepared above is provided. In this embodiment, the biochip prepared in the previous embodiment is positioned face-up, and a sample is applied to the biochip manually or automatically. After the sample reacts with the bioprobes, the results are interpreted visually or by a machine.
[0058] Through the above-mentioned technical features, the biochip manufacturing method of the present application can fix the reaction piece by a simple fixing part, and can be set (placed) by adsorbing the back of the reaction piece during manufacturing, avoiding the damage of the biological probe that may be caused by adsorption on the front side in the traditional process. The biochip prepared is low-cost, and reaction pieces with different reaction components can be set according to needs, thereby further saving costs. For example, 2 or more reaction pieces with different biological probes (and reaction components) can be prepared and placed on the same biochip. In the detection method using the biochip disclosed in the present application, the experimental process is easy to operate and the results are easy to interpret, which is easier to use than the conventional detection method. In addition, since the biochip of the present application is of moderate size, only part of the reaction piece can be used according to needs, which can also save costs.
[0059] The above description is for illustrative purposes only and is not intended to be limiting. Any equivalent modifications or changes made thereto without departing from the spirit and scope of this application shall be included within the scope defined by the claims.
[0060] Explanation of symbols
[0061] 1: Substrate
[0062] 2: Observation hole
[0063] 3,3a: Fixing parts
[0064] 4: Reaction piece
[0065] 5: Fixed part
[0066] S1~S5:Steps
Claims
1. A method for manufacturing a biochip, characterized in that: Manufacture a substrate; Generate a plurality of observation holes penetrating the substrate on the substrate; Arrange a plurality of fixing members around each of the plurality of observation holes on the bottom surface of the substrate; Dispose a plurality of reaction chips respectively corresponding to the plurality of observation holes on one side of the bottom surface; and Fix the plurality of reaction chips respectively through the plurality of fixing members.
2. The manufacturing method according to claim 1, characterized in that, Further include deforming the plurality of fixing members to fix the plurality of reaction chips.
3. The manufacturing method according to claim 2, characterized in that, The deformation is carried out by heating and melting the plurality of fixing members.
4. The manufacturing method according to claim 1, characterized in that, Further include arranging reaction components on a reaction master slice and cutting the reaction master slice into a plurality of reaction chips.
5. The manufacturing method according to claim 1, characterized in that, Each of the reaction chips disposed on the bottom surface includes different reaction components.
6. The manufacturing method according to claim 5, characterized in that, The minimum distance between the centers of each of the reaction chips is 1 to 10 mm.
7. The manufacturing method according to claim 1, characterized in that, The fixing member is an L-shaped column, a cylinder, a polygonal prism, a cone, a cylinder, a sphere, a polyhedron, a ring or a combination thereof.
8. A biochip is manufactured by the method for manufacturing a biochip according to any one of claims 1 to 7.
9. The biochip according to claim 8, characterized in that, Each of the observation holes is surrounded by at least one corresponding fixing member.
10. The biochip according to claim 8, characterized in that, The substrate is made of silicon, glass, a polymer material or ceramics.
11. The biochip according to claim 8, wherein The fixing member is made of plastic, silica gel or rubber.
12. A method for detecting a sample using the biochip according to any one of claims 8 to 11, characterized in that: Add the sample to the reaction chip through the observation hole; and Observe the result from the observation hole.