Gene chip and gene detection device

Abstract

The application discloses a gene chip and a gene chip detection device. The gene chip comprises a substrate and a positioning device fixed to the upper surface of the substrate; the positioning device is provided with a containing cavity used for containing microbeads; the containing cavity is opened on the surface of the positioning device away from the substrate; and the area of the containing cavity is gradually narrowed in the direction of the height of the positioning device towards the upper surface of the substrate. The gene detection device comprises the gene chip and the microbeads covered with fluorescent probes, and the microbeads are clamped in the containing cavity. The gene chip and the gene chip detection device of the application reduce the reflection of the surface of the containing cavity of the substrate to excitation light, and at the same time, the exposed area of the microbeads is increased. In the process of using the gene chip, the proportion of the fluorescence excited by the fluorescent probes on the microbeads in the light received by the detection device is increased, so that the accuracy of the gene chip detection is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of designing a gene chip and a gene detection device, in particular to the field of optical design of the gene detection device. BACKGROUND

[0002] The gene detection device comprises a gene chip, microbeads covered with fluorescent probes, an emitting device and a detection device. The emitting device emits excitation light to excite the fluorescent probes to emit fluorescence. The detection device receives the fluorescence and detects according to the received fluorescence. However, in this process, the gene chip also reflects the excitation light. The excitation light entering the detection device will affect the detection accuracy. In order to accommodate the microbeads, the gene chip usually has holes on the surface of the gene chip for accommodating the microbeads. The surface of the gene chip without the holes is perpendicular to the incident excitation light and reflects the excitation light, so that the reflected excitation light enters the detection device in the direction parallel to the incident excitation light. The detection device receives excitation light with high intensity and high reflectivity, which affects the detection accuracy. SUMMARY

[0003] The gene chip and the gene detection device provided by the embodiments of the present application can improve the accuracy of gene chip detection.

[0004] According to a first aspect of the embodiments of the present application, a gene chip is provided, which comprises a substrate and a positioning device fixed on the upper surface of the substrate; the positioning device is provided with a containing cavity for accommodating microbeads.

[0005] The containing cavity is arranged on the surface of the positioning device away from the substrate; along the height direction of the positioning device, the area of the containing cavity gradually narrows towards the upper surface of the substrate.

[0006] Preferably, the positioning device comprises a plurality of positioning blocks arranged at intervals, and the positioning blocks surround the containing cavity.

[0007] Preferably, the positioning block comprises a plurality of positioning portions, and the positioning portions are at least part of the side wall surface of the positioning block. At least one positioning portion of each positioning block is directed towards the containing cavity.

[0008] Preferably, along the height direction of the positioning device, the positioning portion is away from the center line of the containing cavity towards the upper surface of the substrate.

[0009] Preferably, two adjacent positioning devices can share at least one positioning block.

[0010] Preferably, the positioning device comprises six positioning blocks, and the positioning blocks are evenly distributed along the center line of the containing cavity in the circumferential direction.

[0011] Two adjacent positioning devices share two adjacent positioning blocks.

[0012] Preferably, the positioning block comprises a positioning part as at least part of the side wall surface of the positioning block; each positioning block comprises three uniformly distributed positioning parts.

[0013] The three positioning parts of each positioning block respectively face the accommodating cavity of the positioning device and the accommodating cavities of the other two positioning devices adjacent to the positioning device.

[0014] Preferably, the area of the positioning block gradually narrows away from the upper surface of the substrate in the height direction of the positioning device.

[0015] Preferably, the gene chip is made of transparent material.

[0016] According to a second aspect of the embodiments of the present application, a gene detection device is provided, which comprises the above-mentioned gene chip and the microbeads coated with fluorescent material.

[0017] The microbeads are fixed in the accommodating cavities.

[0018] Preferably, at least part of the microbeads are made of metal material.

[0019] Preferably, the microbeads comprise a body and a coating layer, the coating layer is coated on the outer surface of the body, the fluorescent material is coated on the outside of the coating layer, and the material of the coating layer is the same as that of the substrate.

[0020] The positive progress effect of the present application is that:

[0021] The gene chip and the gene detection device of the present application reduce the reflection of the surface of the accommodating cavity of the substrate to the excitation light by the setting of the surface of the accommodating cavity with an inclination angle, increase the area of the microbeads exposed, and increase the area of the fluorescent probes that can be excited. In the use of the gene chip, the proportion of the fluorescence of the fluorescent probes on the microbeads excited to the light received by the detection device is increased, and the accuracy of the detection of the gene chip is improved. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 is a sectional structure schematic diagram of the gene detection device of the present application.

[0023] Figure 2 is a sectional structure schematic diagram of the gene chip in the gene detection device of the present application.

[0024] Figure 3 is a sectional structure schematic diagram of the gene chip of another embodiment of the present application.

[0025] Figure 4 is a plan view structural schematic of a gene detection device of the present application.

[0026] Figure 5 is another plan view structural schematic of a gene chip in the gene detection device of the present application.

[0027] Figure 6 is a sectional view structural schematic of a microbead of the present application.

[0028] BRIEF DESCRIPTION OF DRAWINGS

[0029] height direction H

[0030] gene chip 100

[0031] substrate 110

[0032] upper surface 111 of the substrate

[0033] positioning device 120

[0034] accommodating cavity 121

[0035] positioning block 122

[0036] positioning portion 123

[0037] upper surface 124 of the positioning block

[0038] lower surface 125 of the positioning block

[0039] microbead 200

[0040] upper hemisphere 201

[0041] lower hemisphere 202

[0042] body 210

[0043] cladding layer 220

[0044] fluorescent probe 230 DETAILED DESCRIPTION

[0045] The exemplary embodiments will now be described in detail with reference to the drawings. When the following description refers to portions, regions, layers, or panels, etc., which are "on" "above" "under" or "below" other portions, regions, layers or panels, etc., it will be understood that these portions, regions, layers, or panels, etc., can be directly on, directly above, directly under, or indirectly connected to the other portions, regions, layers or panels, etc., by one or more intervening portions, regions, layers or panels, etc. The exemplary embodiments will be described herein with reference to the accompanying drawings, which are schematic and are non-limiting. In particular, the figures can not be to scale and certain features can be shown exaggerated in relation to other features for the purpose of explanation. The same reference indicators will be used throughout the drawings and the following description and reference will be made to the drawings to illustrate preferred embodiments of the present application.

[0046] The terminology used in the present application is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. As used in the present application and the following claims, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates 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.

[0047] It should be understood that the use of "first", "second", and / or like designations in the present application and the following claims are used herein to only to help differentiate one element from another and does not necessarily indicate a physical, chronological or spatial sequence or precedence of one element over another. Similarly, "one" or "a" or like designations does not indicate a quantity of one, but rather indicates the presence of at least one of the referenced item. Unless otherwise indicated, "front", "back", "under", and / or "over" and like designations are used for convenience only and are not intended to be limiting as to position or spatial orientation. "Include" or "comprise" and like terms are meant to be equivalent to "including but not limited to," "comprising but not limited to," and "containing, but not limited to," and are not intended to (and should not be construed to) exclude other elements or steps. "Connected" or "coupled" and like terms are not intended to be limited to a direct connection or coupling but also include indirect connections and couplings.

[0048] Embodiments of the present application will be described in detail below with reference to the drawings. The features of the embodiments and examples described below can be combined with each other, as long as there is no conflict.

[0049] As shown in Figures 1 to 6 , the present application discloses a gene detection device, which comprises a gene chip 100, a microbead 200 located on the gene chip 100 and covered with a fluorescent probe 230, an emitting device and a detection device, wherein the emitting device and the detection device are not shown in the figure. The gene detection device emits excitation light to the gene chip 100 and the microbead 200 located on the gene chip 100, the fluorescent probe 230 is excited and emits fluorescence, and at the same time the gene chip 100 reflects the excitation light. The detection device receives the excitation light and the fluorescence, and detects the fluorescence signal according to the received fluorescence. As shown in Figure 1 and Figure 2 , the gene chip 100 comprises a substrate 110 and a positioning device 120. When the gene chip 100 is used for gene detection, the microbead 200 with the outer surface covered with the fluorescent probe needs to be arranged in the positioning device 120 of the gene chip 100, the gene chip 100 is irradiated with excitation light, the fluorescent probe 230 is excited and emits fluorescence, and the detection device detects the gene according to the fluorescence emitted by the fluorescent probe 230.

[0050] As shown in Figure 1 andFigure 2 As shown, the positioning device 120 is fixed on the upper surface 111 of the substrate, and the positioning device 120 is provided with a receiving cavity 121, the receiving cavity 121 is used to receive the microbead 200 with the outer surface covered with the fluorescent probe, and the receiving cavity 121 can only receive one microbead 200. In this embodiment, as shown in FIG. Figure 1 As shown, the microbead 200 is positioned by abutting against the substrate 110 and being located inside the accommodating cavity 121. Of course, the microbead 200 can also be positioned directly by abutting the outer surface of the microbead 200 against the surface of the accommodating cavity 121. The accommodating cavity 121 is opened on the surface of the positioning device 120 away from the substrate 110; along the height direction H of the positioning device 120 toward the upper surface 111 of the substrate, the area of ​​the accommodating cavity 121 gradually narrows. With such an arrangement, the surface of the accommodating cavity 121 is an inclined slope. When the gene chip 100 is irradiated with excitation light, the fluorescent probe is excited and emits fluorescence, and the positioning device 120 also reflects the excitation light. The detection device will not only receive the fluorescence but also the excitation light reflected by the positioning device 120. As shown Figure 2 As shown, by setting the inclined surface, the excitation light reflected by the positioning device 120 is reduced from entering the detection device in the direction parallel to the incident excitation light, and the excitation light received by the detection device is reduced. Figure 2 As shown, located Figure 2 The arrows in the middle represent two incident excitation lights. The excitation lights are reflected on the inclined surface so that the reflected excitation lights do not enter the detection device in a direction parallel to the incident excitation light. Figure 1 As shown, located Figure 1 The left-facing arrow in the middle represents incident excitation light. The inclined surface of the positioning device 120 faces the microbead 200, and the excitation light reflected by the inclined surface can directly hit the surface of the microbead 200 coated with the fluorescent probe, thereby increasing the intensity of the fluorescent probe being excited and emitting fluorescence. Simultaneously, the area of ​​the accommodating cavity 121 gradually narrows from the surface closest to the substrate 110, so that the exposed area of ​​the microbead 200 is greater than 50 percent of the entire outer surface area of ​​the microbead 200. That is, along the vertical direction, the microbead 200 is divided into an upper hemisphere 201 and a lower hemisphere 202. When the surface of the accommodating cavity 121 is vertical, the exposed surface of the microbead 200 is only the surface of the upper hemisphere 201. When the surface of the accommodating cavity 121 is inclined, widening away from the substrate 110, part of the surface of the lower hemisphere 202 of the microbead 200 is also exposed. As the exposed area of ​​the microbead 200 increases, the area receiving the excitation light increases, thereby increasing the probability of excitation of the fluorescent probe and enhancing the intensity of the excited fluorescence. The inclined surface of the accommodating cavity 121 can also reflect part of the excitation light to the surface of the lower hemisphere 202, thereby increasing the area of ​​the microbead 200 receiving the excitation light. In addition, the size of the accommodating cavity 121 can be slightly larger than the size of the microbead 200, such as Figure 1 As shown, located Figure 1The middle right arrow indicates the fluorescence emitted by the fluorescent probe after being excited. The fluorescence can be scattered through the gap between the accommodating cavity 121 and the microbead 200, so that the excited fluorescence of the lower hemisphere 202 can enter the detection device through scattering, thereby enhancing the intensity of the excited fluorescence. Therefore, by arranging the inclined surface, the proportion of the fluorescence received by the detection device can be improved, and the detection accuracy is improved. At the same time, the inclined surface is beneficial to the smooth falling of the microbead 200 into the accommodating cavity 121 for fixation.

[0051] As shown in Figures 1 to 6 , the positioning device 120 includes a plurality of positioning blocks 122 arranged at intervals, and the positioning blocks 122 surround the accommodating cavity 121. By arranging the plurality of positioning blocks 122 at intervals, the flow of the liquid carrying the microbead 200 is facilitated. Of course, the positioning device 120 can also be composed of only one positioning block 122 with a groove on the upper surface. As shown in Figure 4 and Figure 5 , in this embodiment, the number of positioning blocks 122 is six. Of course, in other embodiments, the number of positioning blocks 122 can be two to five, seven or more than seven.

[0052] As shown in Figure 1 and Figure 2 , the positioning block 122 includes a plurality of positioning portions 123, which are at least part of the side surface of the positioning block 122, and at least one positioning portion 123 of each positioning block 122 faces the accommodating cavity 121. By arranging the positioning portion 123, the plurality of positioning portions 123 can form the surface of the accommodating cavity 121, providing a space for accommodating the microbead 200. Among them, along the height direction H of the positioning device 120, the positioning portion 123 is away from the center line of the accommodating cavity 121 towards the direction of the upper surface 111 of the substrate, so as to form the accommodating cavity 121 which gradually narrows in area from the direction close to the upper surface 111 of the substrate. Preferably, along the height direction of the positioning device 120, the area of the positioning block 122 gradually narrows away from the upper surface 111 of the substrate. When the area of the upper surface 124 of the positioning block is smaller than the area of the lower surface 125 of the positioning block, the area of the upper surface 124 of the positioning block arranged vertically to the excitation light is reduced, which can reduce the reflected excitation light parallel to the incident excitation light into the detection device, thereby reducing the background stray light.

[0053] Preferably, two adjacent positioning devices 120 can share at least one positioning block 122. By such arrangement, the utilization rate of the positioning block 122 can be improved, so that a gene chip 100 of the same size with the same number of positioning blocks 122 can accommodate more microbeads 200. As shown in Figure 4 and Figure 5As shown, in the embodiment, each positioning device 120 includes six positioning blocks 122. The positioning blocks 122 are evenly distributed along the center line of the accommodating cavity 121; the six positioning blocks 122 achieve a hexagonal arrangement, which ensures that the microbeads 200 are arranged in the most dense manner in the same plane, thereby increasing the utilization rate of the substrate 110. At this time, two adjacent positioning devices 120 share two adjacent positioning blocks 122. As shown in FIG. 2B, the positioning blocks 122 are arranged in the same plane, and the microbeads 200 are arranged in the most dense manner in the same plane. Figure 4 As shown, the dashed line outlines one gene chip. As shown in FIG. 2C, the dashed line outlines six adjacent gene chips, wherein any two adjacent gene chips share two positioning blocks. Figure 5 As shown, the dashed line outlines one gene chip. As shown in FIG. 2C, the dashed line outlines six adjacent gene chips, wherein any two adjacent gene chips share two positioning blocks. Figure 4 As shown, the dashed line outlines one gene chip. As shown in FIG. 2C, the dashed line outlines six adjacent gene chips, wherein any two adjacent gene chips share two positioning blocks. Figure 1 As shown in FIG. 2D, the three positioning portions 123 of the positioning block 122 are evenly distributed, and the three positioning portions 123 of the positioning block 122 are respectively directed to the accommodating cavity 121 of the positioning device 120 and the accommodating cavities 121 of the other two positioning devices 120 adjacent to the positioning device 120. Figure 2 As shown in FIG. 2D, the three positioning portions 123 of the positioning block 122 are evenly distributed, and the three positioning portions 123 of the positioning block 122 are respectively directed to the accommodating cavity 121 of the positioning device 120 and the accommodating cavities 121 of the other two positioning devices 120 adjacent to the positioning device 120. Figure 4 As shown in FIG. 2D, the three positioning portions 123 of the positioning block 122 are evenly distributed, and the three positioning portions 123 of the positioning block 122 are respectively directed to the accommodating cavity 121 of the positioning device 120 and the accommodating cavities 121 of the other two positioning devices 120 adjacent to the positioning device 120. Figure 5 As shown in FIG. 2D, the three positioning portions 123 of the positioning block 122 are evenly distributed, and the three positioning portions 123 of the positioning block 122 are respectively directed to the accommodating cavity 121 of the positioning device 120 and the accommodating cavities 121 of the other two positioning devices 120 adjacent to the positioning device 120. Figure 3 As shown in FIG. 2D, the three positioning portions 123 of the positioning block 122 are evenly distributed, and the three positioning portions 123 of the positioning block 122 are respectively directed to the accommodating cavity 121 of the positioning device 120 and the accommodating cavities 121 of the other two positioning devices 120 adjacent to the positioning device 120. Figure 1As shown, in the present embodiment, the height h of the positioning block 122 is 400 nm, the bottom surface diameter D1 of the positioning block is 500 nm, and the minimum distance L between two adjacent positioning blocks is 1300 nm, and the diameter D2 of the microbead 200 is 1300 nm. Among them, in order to better reflect the optical path of the incident light, the optical path of the fluorescence, and the structure of the positioning device and the substrate, the size ratio between the positioning block 122, the microbead 200 and the substrate 110 is not drawn according to the true proportion. Of course, the height, the bottom surface diameter of the positioning block, and the minimum distance between two adjacent positioning blocks 122 can be adjusted according to the size of the corresponding microbead 200.

[0054] Preferably, the gene chip 100 is made of transparent material, that is, the substrate 110 and the positioning device 120 are made of transparent material, through such a setting, the reflectivity of the positioning device 120 and the substrate 110 to the excitation light is reduced. At the same time, the transparent material needs to be selected from non-toxic materials to ensure the safety of the user and the fluorescence probe is not contaminated. Preferably, the material of the gene chip 100 can be selected from glass or resin, of course, the material of the gene chip 100 can also be selected from other transparent and non-toxic materials that are easy to nanoimprint, which reduces the reflectivity of the positioning device 120 and the substrate 110 to the excitation light, and facilitates the processing and molding of the gene chip 100. Among them, in the present embodiment, the substrate 110 and the positioning device 120 are separable components, such a setting makes it convenient to clean when the gene chip 100 is repeatedly used, so that the structure of the last experiment does not affect the next experiment, of course, the substrate 110 and the positioning device 120 can be integrally formed, to prevent the loss of part of the positioning device 120 or the substrate 110.

[0055] As Figure 6As shown, at least part of the microbead 200 is made of a metal material. The body 210 of the metal material can form a localized surface plasmon resonance, which is experimentally shown to be beneficial to the excitation of the fluorescent probe 230. Preferably, the microbead 200 comprises the body 210 and a coating layer 220, which coats the outer surface of the body 210. The body 210 is made of a metal material. With such an arrangement, simulation results show that the Purcell factor is 1.18, according to the definition formula of the Purcell factor, the fluorescence intensity of the fluorescent probe 230 is enhanced by 18% in the structure of the present application, and the fluorescence intensity is enhanced. The material of the body 210 can be gold, silver, copper and aluminum, and of course the material of the body 210 can also be other metal materials. The fluorescent probe 230 is arranged outside the coating layer 220, and the material of the coating layer 220 is the same as that of the substrate 110. When the material of the coating layer 220 is the same as that of the substrate 110, there is a van der Waals force between the microbead 200 and the surface of the holding cavity 121 or the substrate 110, which is beneficial to the fixation of the microbead 200. The coating layer 220 can also separate the metal material microbead 200 from the direct contact with the fluorescent probe 230, so as to avoid the quenching phenomenon of the fluorescent substance of the fluorescent probe 230. At the same time, the material of the coating layer 220 is the same as that of the gene chip 100, which is a non-toxic material, and the amino or mercapto which can form a covalent bond with DNA can be synthesized on the surface of the coating layer 220, and the gene probe with fluorescent substance can be implanted to form the fluorescent probe 230. In the embodiment, the thickness of the coating layer is 150 nm, and of course the thickness of the coating layer can be adjusted according to the diameter of the body or actual needs. The structure of the coating layer and the body is clearly shown, and the size of the coating layer and the body is not drawn according to the true proportion.

[0056] The above description is only the preferred embodiment of the present application, and does not limit the present application in any form. Although the present application has been disclosed as the above preferred embodiment, it is not intended to limit the present application. Any skilled person in the art can make some changes or modifications to the above disclosed technical content without departing from the scope of the technical solution of the present application, and any simple modification, equivalent change and modification of the above embodiment according to the technical essence of the present application are still within the scope of the technical solution of the present application.

[0057] The content disclosed in the patent document contains copyrighted material. The copyright is owned by the copyright owner. The copyright owner does not object to the copying of the patent document or the patent disclosure in the official records and archives of the patent and trademark office.

Claims

1. A gene chip, characterized in that The gene chip includes a substrate and a positioning device fixed to the upper surface of the substrate; the positioning device is provided with a receiving cavity, the receiving cavity is used to receive microbeads, the surface of the microbeads is covered with fluorescent probes, the fluorescent probes are used to be excited and emit fluorescence under the irradiation of excitation light, and the light cannot penetrate the microbeads; The accommodating cavity is opened on the surface of the positioning device away from the substrate; along the height direction of the positioning device toward the upper surface of the substrate, the area of ​​the accommodating cavity gradually narrows; The positioning device includes six positioning blocks arranged at intervals, the positioning blocks are evenly distributed along the circumference of the center line of the accommodating cavity, and the positioning blocks surround the accommodating cavity; Two adjacent positioning devices share two adjacent positioning blocks; The positioning block includes a positioning portion, which serves as at least a portion of a side wall surface of the positioning block; each positioning block includes three evenly distributed positioning portions; The three positioning parts of each positioning block are respectively oriented toward the accommodating cavity of a positioning device and the accommodating cavities of the other two positioning devices adjacent to the positioning device; The area of ​​the positioning block gradually narrows along the height direction of the positioning device toward a direction away from the upper surface of the substrate.

2. The gene chip according to claim 1, wherein Along the height direction of the positioning device toward the upper surface of the substrate, the positioning portion is away from the center line of the accommodating cavity.

3. The gene chip according to claim 1 or 2, wherein The gene chip is made of transparent material.

4. A gene detection device, characterized in that: The gene detection device comprises the gene chip according to any one of claims 1 to 3 and the microbeads coated with fluorescent material; The micro beads are fixed in the accommodating cavity.

5. The gene detection device according to claim 4, wherein At least a portion of the microbeads is made of a metallic material.

6. The gene detection device according to claim 5, wherein The microbeads include a body and a coating layer, wherein the coating layer coats the outer surface of the body, and a fluorescent material is coated on the outer side of the coating layer. The material of the coating layer is the same as that of the substrate.

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