Gene chip hybridization box, gene chip set and gene chip hybridization method

Abstract

The present application relates to a gene chip hybridization box, a gene chip set, and a gene chip hybridization method. The gene chip hybridization box includes: a substrate having a gene chip placement area, a first surface of a gene chip capable of being bonded to the gene chip placement area; a cover plate having a through hole, the through hole being opposite to the gene chip placement area; a spacer disposed between the substrate and the cover plate, the spacer being used to create a predetermined gap between the substrate and the cover plate, and the spacer having an opening in an area facing at least one gene chip placement area, thereby defining a gene chip accommodation space between the substrate and the cover plate; the predetermined gap being set so that when the gene chip is fixed in the gene chip placement area, a second surface of the gene chip, which is opposite to the first surface and includes a probe area, is spaced from a surface of the cover plate facing the second surface by 0.05 mm to 0.15 mm, so that when a hybridization solution is injected through the through hole of the cover plate, the hybridization solution can spread over the second surface by capillary action.

Description

Technical Field

[0001] The present application relates to the field of biotechnology, and in particular to a gene chip hybridization box, a gene chip set and a gene chip hybridization method. Background Art

[0002] Gene chips are an emerging biotechnology used for DNA sequencing, identification of gene expression profiles, and detection and analysis of gene mutants. The basic principle of gene chips is to design probes based on known nucleotide sequences, hybridize them with labeled target nucleotide sequences, and perform qualitative and quantitative analysis through signal detection. Gene chips integrate a large number of molecular recognition probes on the surface of a tiny substrate (silicon wafer, glass wafer, etc.). These probes hybridize with labeled samples, and the genetic information of the sample is obtained through detection and analysis of the hybridization signals. This allows for the simultaneous analysis of numerous genes, enabling detection and analysis of large amounts of information.

[0003] In situ synthesis microarrays, a type of DNA chip developed by Affymetrix in the United States, are made by synthesizing oligonucleotides in situ at specific locations on the chip. In situ synthesis methods include photodeprotection, inkjet synthesis, photoacid synthesis, electrospray synthesis, virtual masking, and molecular stamping. In situ synthesis microarrays are high-density gene chips that integrate a large number of densely packed gene probes, enabling simultaneous analysis of numerous genes and rapid acquisition of genetic information.

[0004] Liquid-phase hybridization, the selective binding between target sample nucleic acid molecules and chip probes, is a form of solid-liquid hybridization. Hybridization involves hybridizing a labeled sample in the liquid phase with a probe of known sequence immobilized on the solid surface of the chip. This allows for the simultaneous detection of multiple gene sequences. Hybridization efficiency is influenced by numerous factors, including liquid salt concentration, temperature, reaction time, and DNA secondary structure. After hybridization, the higher the degree of match between the sample and probe, the greater the thermodynamic stability and the stronger the generated fluorescence signal. Sample information is obtained by scanning, collecting, comparing, and analyzing the hybridization signals (including location, intensity, and color) with software.

[0005] When the hybridization solution is brought into contact with the surface of the chip containing the probe area for hybridization, the chip needs to be immersed in the hybridization solution within a certain space to ensure that the hybridization solution is in full contact with the chip surface. Therefore, the hybridization solution in this space (such as a deep well) needs to reach a specified volume. In order to achieve the detection effect, various reagents need to reach a specified concentration in the hybridization solution, so the amount of reagents used is proportional to the specified volume of the hybridization solution. Various expensive reagents need to be used in the detection, so it is necessary to reduce the specified volume of the hybridization solution to effectively reduce the cost of a single test.

[0006] However, when the amount of hybridization solution is reduced, there is a risk that the hybridization solution fails to fully cover the chip surface and thus cannot fully contact and hybridize with the chip surface. Summary of the Invention

[0007] In order to overcome or at least partially alleviate the above-mentioned technical problems, according to a first aspect of the present application, a gene chip hybridization box is provided, comprising: a substrate having at least one gene chip arrangement area, to which a first surface of a gene chip not including a probe area can be bonded; a cover plate, the cover plate being provided with at least one through hole, wherein the through hole is arranged opposite to the gene chip arrangement area; and a spacer arranged between the substrate and the cover plate, the spacer being used to create a predetermined gap between the substrate and the cover plate, and the spacer having an opening in a region facing the at least one gene chip arrangement area, thereby defining a gene chip accommodating space between the substrate and the cover plate; wherein the predetermined gap is arranged such that when the gene chip is fixed in the gene chip arrangement area, a second surface of the gene chip opposite to the first surface and including the probe area is spaced from a surface of the cover plate facing the second surface by 0.05 mm to 0.15 mm, so that when a hybridization solution is injected through the at least one through hole of the cover plate, the hybridization solution can spread over the second surface of the gene chip by capillary action.

[0008] According to one embodiment, the gene chip placement area is provided with a recess for fixing and accommodating the gene chip in a tight-fitting manner.

[0009] According to one embodiment, the substrate is provided with at least two positioning hooks for fixing the gene chip in the gene chip arrangement area.

[0010] According to one embodiment, the substrate and the spacer are formed integrally; or the cover and the spacer are formed integrally.

[0011] According to one embodiment, the gene chip arrangement area is provided with at least one closed flange for fixing and accommodating the gene chip by tight fit, and when the substrate, spacer and cover are fixed to each other, the closed flange abuts the cover, thereby forming a closed space between the closed flange and the cover.

[0012] According to one embodiment, the cover plate is provided with an exhaust hole communicating with the enclosed space at a position opposite to an edge portion of the enclosed space.

[0013] According to one embodiment, the through hole is arranged to be opposite to the center position of the gene chip.

[0014] According to one embodiment, the predetermined interval is set so that when at least one gene chip is fixed on the substrate, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the cover facing the second surface is between 0.08mm and 0.12mm.

[0015] According to one embodiment, the gene chip hybridization box is further provided with at least one clamping device for releasably clamping the base plate, the spacer, and the cover plate together.

[0016] According to one embodiment, a conductive element is provided in the gene chip placement region, and the conductive element is arranged so that when a first surface of the gene chip that does not include the probe region is bonded to the gene chip placement region, the conductive element is electrically connected to the first surface.

[0017] According to one embodiment, the substrate is formed of a conductive material, so that when a first surface of the gene chip that does not include the probe region is bonded to the gene chip placement area, the conductive material is electrically connected to the first surface.

[0018] According to the above embodiment, by setting the interval between the second surface of the gene chip and the surface of the cover plate facing the second surface, the hybridization liquid is spread over (infiltrated) the second surface of the gene chip through capillary action, so that the hybridization liquid is fully in contact with the second surface of the gene chip for hybridization.

[0019] According to a second aspect of the present application, a gene chip hybridization box is provided, comprising: a substrate; a cover plate, the cover plate having at least one gene chip arrangement area, a first surface of the gene chip not including a probe area being capable of being bonded to the gene chip arrangement area, the cover plate being provided with a through hole at a position outside the gene chip arrangement area and adjacent to the gene chip arrangement area; and a spacer arranged between the substrate and the cover plate, the spacer being used to create a predetermined interval between the substrate and the cover plate, and the spacer having an opening in an area facing the at least one gene chip arrangement area, thereby defining a gene chip accommodating space between the substrate and the cover plate; wherein the predetermined interval is arranged so that when the gene chip is fixed in the gene chip arrangement area, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the substrate facing the second surface is between 0.05 mm and 0.15 mm, so that when a hybridization solution is injected through the at least one through hole of the cover plate, the hybridization solution can spread over the second surface of the gene chip by capillary action.

[0020] According to one embodiment, the gene chip placement area is provided with a recess for fixing and accommodating the gene chip in a tight-fitting manner.

[0021] According to one embodiment, the cover plate is provided with at least two positioning hooks for fixing the gene chip in the gene chip arrangement area.

[0022] According to one embodiment, the substrate and the spacer are formed integrally; or the cover and the spacer are formed integrally.

[0023] According to one embodiment, the cover plate is provided with a closed flange surrounding the gene chip arrangement area and the adjacent through holes, and when the substrate, spacer and cover plate are fixed to each other, the closed flange abuts the substrate, thereby forming a closed space between the closed flange and the substrate; or the substrate is provided with a closed flange surrounding the gene chip arrangement area and the adjacent through holes, and when the substrate, spacer and cover plate are fixed to each other, the closed flange abuts the cover plate, thereby forming a closed space between the closed flange and the cover plate.

[0024] According to one embodiment, the cover plate is provided with an exhaust hole communicating with the enclosed space.

[0025] According to one embodiment, the through hole and the exhaust hole are arranged to be opposite to each other along a diagonal line of the gene chip arrangement area.

[0026] According to one embodiment, the predetermined interval is set so that when at least one gene chip is fixed on the substrate, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the cover facing the second surface is between 0.08mm and 0.12mm.

[0027] According to one embodiment, the gene chip hybridization box is further provided with at least one clamping device for releasably clamping the base plate, the spacer, and the cover plate together.

[0028] According to one embodiment, a conductive element is provided in the gene chip placement region, and the conductive element is arranged so that when a first surface of the gene chip that does not include the probe region is bonded to the gene chip placement region, the conductive element is electrically connected to the first surface.

[0029] According to one embodiment, the cover plate is formed of a conductive material, so that when a first surface of the gene chip that does not include the probe region is bonded to the gene chip placement area, the conductive material is electrically connected to the first surface.

[0030] The embodiment according to the second aspect can achieve the same beneficial technical effect as the embodiment according to the first aspect (i.e., the hybridization solution is spread over the second surface of the gene chip by capillary action). In addition, because the through hole for injecting the hybridization solution is located on the side of the gene chip and away from the second surface of the gene chip containing the probes, it is possible to prevent the tip of the pipette from accidentally contacting the probes on the second surface of the gene chip when injecting the hybridization solution, thereby preventing the probes from being damaged.

[0031] According to the third aspect of the present application, a gene chip kit is further provided, comprising: at least one gene chip; and the above-mentioned gene chip hybridization box.

[0032] According to the fourth aspect of the present application, a gene chip hybridization method is also provided, which comprises the following steps: providing the above-mentioned gene chip hybridization box; arranging the gene chip to be hybridized in the gene chip hybridization box; and allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area through capillary action.

[0033] According to one embodiment, the step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area through capillary action includes: placing the gene chip hybridization box so that the substrate of the gene chip hybridization box is located below the cover plate; and injecting the hybridization solution into the gene chip hybridization box through the through hole of the gene chip hybridization box.

[0034] According to one embodiment, the step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area through capillary action includes: placing the gene chip hybridization box so that the cover of the gene chip hybridization box is located below the base plate; and injecting the hybridization solution into the gene chip hybridization box through the through hole of the gene chip hybridization box.

[0035] According to one embodiment, the step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and to cover the second surface of the gene chip containing the probe area through capillary action includes: placing the gene chip hybridization box so that the cover of the gene chip hybridization box is located below the base plate; and at least partially immersing the cover in the hybridization solution so that the hybridization solution covers the second surface of the gene chip containing the probe area through capillary action.

[0036] According to one embodiment, the method further comprises: applying an electric field to the gene chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Below, the preferred embodiments of the present application will be described in detail with reference to the schematic drawings. In the drawings:

[0038] Figure 1A and Figure 1B are respectively a front view and a side view of a substrate according to a first embodiment of the present application;

[0039] Figure 2A and Figure 2B They are respectively a front view and a side sectional view of a cover plate according to a first embodiment of the present application;

[0040] Figure 3A and Figure 3B are respectively a front view and a side sectional view of a spacer according to a first embodiment of the present application;

[0041] Figure 4 is a side cross-sectional view of the substrate, spacer, and cover plate assembled together according to the first embodiment of the present application, wherein a pipette gun is also shown;

[0042] Figure 5A and Figure 5B They are respectively a front view of a substrate and a side cross-sectional view passing through the center line of a gene chip according to a second embodiment of the present application;

[0043] Figure 6 is a side cross-sectional view of a substrate, a spacer, and a cover plate assembled together according to a second embodiment of the present application, wherein a pipette gun is also shown;

[0044] Figure 7A and Figure 7B are respectively a front view and a side view of a substrate according to a third embodiment of the present application;

[0045] Figure 8A and Figure 8B are respectively a front view and a side sectional view of a cover plate according to a third embodiment of the present application; and

[0046] Figure 9 This is a side cross-sectional view of a substrate, a spacer, and a cover plate assembled together according to a third embodiment of the present application, wherein a pipette gun is also shown.

[0047] The accompanying drawings illustrate exemplary embodiments and constitute a part of the specification. Together with the description of the specification, they serve to explain exemplary implementation of the embodiments. The illustrated embodiments are for illustrative purposes only and do not limit the scope of protection of the claims. Throughout the drawings, the same reference numerals refer to the same or similar elements. DETAILED DESCRIPTION

[0048] In this application, unless otherwise specified, the use of terms such as "first" and "second" to describe various elements is not intended to limit the positional relationship, temporal relationship, or importance relationship of these elements. Such terms are simply used to distinguish one element from another. In some examples, the first element and the second element may refer to the same example of the element, while in some cases, based on the context of the description, they may also refer to different examples.

[0049] The terms used in the descriptions of the various examples described in this application are for the purpose of describing specific examples only and are not intended to be limiting. Unless the context clearly indicates otherwise, if the number of elements is not specifically limited, the element may be one or more. In addition, the term "and / or" used in this application encompasses any and all possible combinations of the listed items.

[0050] Next, we will combine Figures 1A to 9 An embodiment of the gene chip hybridization box according to the present application is described in detail.

[0051] Figures 1A to 4 The gene chip hybridization box 100 of the first embodiment of the present application is shown, and the gene chip hybridization box 100 includes: a substrate 110 ( Figure 1A and Figure 1B ), the substrate 110 has a plurality of gene chip placement areas 111, a first surface (not shown) of the gene chip 200 that does not include a probe area can be bonded to the gene chip placement area 111, and the gene chip placement area 111 is provided with a recess 112 for fixing and accommodating the gene chip 200 in a manner such as tight fit; a cover plate 120 ( Figure 2A and Figure 2B ), the cover plate 120 is provided with a plurality of through holes 121, wherein the through holes 121 are arranged to be opposite to the center position of the gene chip arrangement area 111 and thus opposite to the center position of the gene chip 200 (see Figure 4 ); and a spacer 130 ( disposed between the substrate 110 and the cover plate 120 ); Figure 3A and Figure 3B ), the spacer 130 is used to create a predetermined distance d between the substrate 110 and the cover 120 (see Figure 4), and the spacer 130 has an opening 131 in the area facing the gene chip arrangement area 111, thereby defining a gene chip accommodating space 131a between the substrate 110 and the cover plate 120; wherein the predetermined interval d is set so that when the gene chip 200 is fixed in the gene chip arrangement area 111, the second surface 210 of the gene chip 200 opposite to the first surface and including the probe area and the surface of the cover plate 120 facing the second surface 210 are spaced d1 between 0.05 mm and 0.15 mm, preferably between 0.08 mm and 0.12 mm, and for example can be 0.08 mm, 0.09 mm, 0.10 mm, 0.11 mm, etc., so that when the hybridization solution is injected through the through hole 121 of the cover plate 120, the hybridization solution can cover the second surface 210 of the gene chip 200 by capillary action.

[0052] Optionally, the gene chip hybridization box 100 according to the present application is further provided with at least one clamping device (not shown) for releasably clamping the substrate 110, the cover 120, and the spacer 130 together. The clamping device can have any structure known to those skilled in the art and will not be described in detail here.

[0053] exist Figures 1A to 4 In the illustrated embodiment, the gene chip 200 is fixed in the gene chip placement area 111 by tightly fitting with the recess 112. However, it should be understood that the gene chip 200 may be fixed to the gene chip placement area 111 by other means. For example, the substrate 110 is provided with at least two, and preferably at least three, positioning hooks for securing the gene chip 200 in the gene chip placement area 111. The positioning hooks may have the shape and structure of hooks commonly found in the field of electrical connectors, such as the shape and structure of the hooks described and illustrated in CN205985544U.

[0054] exist Figures 1A to 4 In the embodiment shown, the through hole 121 is arranged opposite to the center of the gene chip arrangement area 111 and thus opposite to the center of the gene chip 200 (see Figure 4 ). It should be understood that the through hole 121 can also be arranged to be opposite to the edge position of the gene chip arrangement area 111 and therefore opposite to the edge position of the gene chip 200, and preferably the through hole 121 is arranged to be offset from the probe area of ​​the second surface 210 of the gene chip 200 to avoid the tip of the pipette 400 accidentally touching the probe area of ​​the second surface 210 of the gene chip 200 when the hybridization solution is injected through the through hole 121 by the pipette 400, thereby damaging the probe area.

[0055] exist Figures 1A to 4 In the illustrated embodiment, the substrate 110, the cover 120, and the spacer 130 are shown as three separate components. However, it should be understood that, depending on the application, the substrate 110 and the spacer 130 may be integrally formed; or the cover 120 and the spacer 130 may be integrally formed.

[0056] According to the second embodiment of the present application, as the first embodiment Figure 1A and Figure 1B Variations of the substrate 110 shown, such as Figure 5A and 5B As shown, the gene chip placement area 111 is provided with a closed flange 113 for fixing and accommodating the gene chip 200 by tight fit. When the substrate 110, the spacer 130 and the cover 120 are fixed to each other (see Figure 6 ), the closed flange 113 abuts the cover plate 120, thereby forming a closed space 113a between the closed flange 113 and the cover plate 120. Furthermore, to prevent air in the closed space 113a from being smoothly exhausted and thereby hindering the smooth flow of the hybridization liquid, the cover plate 120 is provided with an exhaust hole 122 at a position opposite to the edge of the closed space 113a, which is in communication with the closed space 113a.

[0057] The cover plate 120 and the spacer 130 according to the second embodiment of the present application are the same as the cover plate 120 and the spacer 130 of the first embodiment, and thus will not be described in detail herein.

[0058] In the first and second embodiments described above, a conductive element (not shown) is provided in the gene chip placement region 111 of the substrate 110. The conductive element is configured such that when the first surface of the gene chip 200, excluding the probe region, is bonded to the gene chip placement region 111, the conductive element electrically connects to the first surface, thereby applying an electric field to the gene chip 200. This addresses the issue of the gene chip 200 being negatively charged and thus not easily hybridizing with DNA. Furthermore, the conductive element can be used to adjust the potential of the gene chip to better meet detection requirements.

[0059] Alternatively, in the above-mentioned first and second embodiments, the substrate 110 can be made of a conductive material, so that when the first surface of the gene chip 200 that does not include the probe area is bonded to the gene chip arrangement area 111, the conductive material is electrically connected to the first surface. This can also solve the problem that the gene chip 200 is not easy to hybridize DNA due to its negative charge, and can adjust the potential of the gene chip to better meet the detection needs.

[0060] The conductive element and conductive material may include any conductive material, such as iron, copper, aluminum, tin, silver, platinum, gold, and any combination and / or alloy thereof, and may be, for example, stainless steel.

[0061] According to the above-mentioned first and second embodiments, by setting the interval between the second surface 210 of the gene chip 200 and the surface of the cover plate 120 facing the second surface 210, the hybridization liquid is spread over (infiltrated) the second surface 210 of the gene chip 200 through capillary action, so that the hybridization liquid is fully in contact with the second surface 210 of the gene chip 200 for hybridization, and bubbles in the hybridization liquid on the second surface 210 can be avoided.

[0062] According to the second embodiment of the present application, compared with the first embodiment, since a closed space 113a is formed by providing a closed flange 113, while ensuring that the hybridization liquid can cover the entire second surface 210 of the gene chip 200 through capillary action, unnecessary hybridization liquid filling space is further reduced, thereby further reducing the amount of hybridization liquid used.

[0063] In the first and second embodiments described above, the gene chip 200 is placed in the gene chip placement area 111 on the substrate 110, and the through hole for injecting the hybridization solution is provided on the cover plate 120. As a feasible embodiment (hereinafter referred to as the "third embodiment"), the gene chip 200 and the through hole for injecting the hybridization solution may also be provided on the cover plate.

[0064] like Figure 3A and Figure 3B as well as Figures 7A to 9 As shown, the gene chip hybridization box 300 according to the third embodiment of the present application includes: a substrate 310 ( Figure 7A and Figure 7B ), the substrate 310 is a flat plate; the cover 320 ( Figure 8A and Figure 8B ), the cover plate 320 has a plurality of gene chip placement areas 321, the first surface of the gene chip 200 not including the probe area is bonded to the gene chip placement area 321, the cover plate 320 is provided with through holes 322 outside and adjacent to the gene chip placement area 321; and a spacer 130 ( Figure 3A and Figure 3B ), the spacer 130 is used to create a predetermined gap d between the substrate 310 and the cover 320 ( Figure 9), and the spacer 130 has an opening 131 in the area facing the gene chip arrangement area 321, thereby defining a gene chip accommodating space 131a between the substrate 310 and the cover plate 320; wherein the predetermined interval d is set so that when the gene chip 200 is fixed in the gene chip arrangement area 321, the interval d1 between the second surface 210 of the gene chip 200 opposite to the first surface and including the probe area and the surface of the substrate 310 facing the second surface 210 is between 0.05mm and 0.15mm, preferably between 0.08mm and 0.12mm, and for example can be 0.08mm, 0.09mm, 0.10mm, 0.11mm, etc., so that when the hybridization solution is injected through the through hole 322 of the cover plate 320, the hybridization solution can cover the second surface 210 of the gene chip 200 by capillary action.

[0065] Similar to the first and second embodiments described above, the gene chip placement area 321 may be provided with a recess for tightly securing and accommodating the gene chip 200, and / or the cover plate 320 may be provided with at least two positioning hooks for securing the gene chip 200 within the gene chip placement area 321. The base plate 310 and the spacer 130 may be integrally formed; alternatively, the cover plate 320 and the spacer 130 may be integrally formed. Furthermore, the gene chip hybridization box 300 may optionally be provided with at least one clamping device for releasably clamping the base plate 310, the spacer 130, and the cover plate 320 together.

[0066] Furthermore, similar to the second embodiment, in the third embodiment, the cover plate 320 may be provided with a closed flange (not shown) surrounding the gene chip placement area 321 and the adjacent through-holes 322. When the substrate 310, the spacer 130, and the cover plate 320 are fixed to each other, the closed flange abuts the substrate 310, thereby forming a closed space between the closed flange and the substrate 310. Alternatively, the substrate 310 may be provided with a closed flange (not shown) surrounding the gene chip placement area 321 and the adjacent through-holes 322. When the substrate 310, the spacer 130, and the cover plate 320 are fixed to each other, the closed flange abuts the cover plate 320, thereby forming a closed space between the closed flange and the cover plate 320. In both cases, the cover plate 320 is provided with a vent (not shown) communicating with the closed space, and preferably, the through-holes 322 and the vent are arranged opposite each other along a diagonal line of the gene chip placement area 321.

[0067] Similar to the first and second embodiments described above, in the third embodiment, a conductive element (not shown) is provided in the gene chip placement area 321 of the cover plate 320. This conductive element is configured such that when the first surface of the gene chip 200, excluding the probe region, is bonded to the gene chip placement area 111, the conductive element electrically connects to the first surface, thereby applying an electric field to the gene chip 200. This addresses the issue of the gene chip 200 being negatively charged and thus not easily hybridizing with DNA. Furthermore, the conductive element allows the potential of the gene chip to be adjusted to better meet detection requirements.

[0068] Alternatively, in the third embodiment, the cover plate 320 can be made of a conductive material. When the first surface of the gene chip 200, excluding the probe region, is bonded to the gene chip placement area 111, the conductive material is electrically connected to the first surface. This can also address the issue of the gene chip 200 being negatively charged and thus less susceptible to DNA hybridization, and can also adjust the potential of the gene chip to better meet detection requirements. Furthermore, the conductive element can be used to adjust the potential of the gene chip to better meet detection requirements.

[0069] Similarly, the conductive elements and conductive materials mentioned above may include any conductive material, such as iron, copper, aluminum, tin, silver, platinum, gold, and any combination and / or alloy thereof, and may be, for example, stainless steel.

[0070] The third embodiment of the present application can achieve the same beneficial technical effects as the first and second embodiments (i.e., the hybridization solution is spread over the second surface of the gene chip through capillary action). In addition, because the through hole 322 for injecting the hybridization solution is located on the side of the gene chip 200 and away from the second surface 210 of the gene chip 200 containing the probes, it can prevent the tip of the pipette 400 from accidentally contacting the probes on the second surface 210 of the gene chip 200 when injecting the hybridization solution, thereby preventing the probes from being damaged.

[0071] It should be noted that, since the gene chip hybridization boxes 100 and 300 according to the first embodiment, the second embodiment and the third embodiment of the present application distribute the hybridization solution on the second surface 210 of the gene chip 200 by capillary action, the gene chip hybridization boxes 100 and 300 according to the first embodiment, the second embodiment and the third embodiment of the present application can be respectively Figure 4 、 Figure 6 and Figure 9 The state shown in FIG (i.e. the substrate is under the cover) can also be used Figure 4 、 Figure 6 and Figure 9 Use it in the state after rotating 180 degrees (i.e. the cover is under the base).

[0072] When the gene chip hybridization cassettes 100 and 300 according to the first embodiment, the second embodiment and the third embodiment of the present application are Figure 4 、 Figure 6 and Figure 9 When used in the 180-degree rotated position (i.e., the cover plate is below the base plate), the hybridization solution enters from the bottom of the gene chip hybridization box and is distributed on the second surface 210 of the gene chip 200 by capillary action. In this case, the hybridization solution can be injected using a pipette 400, or the cover plate can be immersed in the hybridization solution, allowing the hybridization solution to be automatically drawn into the gap between the second surface 210 of the gene chip 200 and the cover plate 120 (for the first and second embodiments) or into the gap between the second surface 210 of the gene chip 200 and the base plate 310 (for the third embodiment), thereby being distributed on the second surface 210 of the gene chip 200.

[0073] According to the present application, a gene chip set is further provided, comprising: at least one gene chip 200; and the gene chip hybridization box according to the first embodiment, the second embodiment, and / or the third embodiment.

[0074] According to the present application, a gene chip hybridization method is also provided, which comprises the following steps: providing a gene chip hybridization box according to the first, second or third embodiment; arranging the gene chip to be hybridized in the gene chip hybridization box; and allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area through capillary action.

[0075] According to a feasible embodiment, the step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area by capillary action includes: placing the gene chip hybridization box so that the substrate of the gene chip hybridization box is located below the cover plate (i.e. Figure 4 、 Figure 6 and Figure 9 and injecting the hybridization solution into the gene chip hybridization box through the through hole of the gene chip hybridization box.

[0076] According to a feasible embodiment, the step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area by capillary action includes: placing the gene chip hybridization box so that the cover of the gene chip hybridization box is located below the base plate (i.e. Figure 4 、 Figure 6 and Figure 9and injecting the hybridization solution into the gene chip hybridization box through the through hole of the gene chip hybridization box.

[0077] According to a feasible embodiment, the step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area by capillary action includes: placing the gene chip hybridization box so that the cover of the gene chip hybridization box is located below the base plate (i.e. Figure 4 、 Figure 6 and Figure 9 The cover plate is at least partially immersed in the hybridization solution, so that the hybridization solution is spread over the second surface of the gene chip containing the probe area by capillary action. In this way, the hybridization solution can be injected into all gene chips in the gene chip box at once, significantly improving operational efficiency.

[0078] According to a feasible embodiment, the method may further include applying an electric field to the gene chip. By applying the electric field to the gene chip, the negative charge carried by the gene chip 200 can be eliminated, thereby promoting DNA hybridization. In addition, the potential of the gene chip can also be adjusted to better meet detection needs.

[0079] Although the embodiments of the present application have been described with reference to the accompanying drawings, it should be understood that the scope of the present application is not limited by these embodiments or examples, but is limited only by the claims and their equivalents after authorization. Various elements in the embodiments or examples may be omitted or replaced by their equivalents. It should also be understood that as technology evolves, many of the elements described herein may be replaced by equivalent elements that appear after the present application.

Claims

1. A gene chip hybridization box, characterized in that: The gene chip hybridization box comprises: A substrate having at least one gene chip placement area, to which a first surface of the gene chip excluding the probe area can be bonded, wherein the gene chip placement area is provided with a recess for fixing and accommodating the gene chip in a tight fit manner; a cover plate, the cover plate being provided with at least one through hole, wherein the through hole is arranged opposite to the gene chip arrangement area; and a spacer disposed between the substrate and the cover plate, the spacer being used to create a predetermined gap between the substrate and the cover plate, and the spacer having an opening in a region facing the at least one gene chip arrangement region, thereby defining a gene chip accommodating space between the substrate and the cover plate; In which, the predetermined interval is set so that when the gene chip is fixed in the gene chip layout area, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the cover plate facing the second surface is between 0.05mm and 0.15mm, so that when the hybridization liquid is injected through at least one through hole of the cover plate, the hybridization liquid can spread over the second surface of the gene chip by capillary action, and the gene chip layout area is provided with at least one closed flange for fixing and accommodating the gene chip by tight fit, and when the substrate, spacer and cover plate are fixed to each other, the closed flange abuts the cover plate, thereby forming a closed space between the closed flange and the cover plate.

2. The gene chip hybridization cassette according to claim 1, characterized in that The substrate is provided with at least two positioning hooks for fixing the gene chip in the gene chip arrangement area.

3. The gene chip hybridization cassette according to claim 1, characterized in that The substrate and the spacer are formed integrally; or the cover and the spacer are formed integrally.

4. The gene chip hybridization cassette according to claim 1, characterized in that The cover plate is provided with an exhaust hole communicating with the closed space at a position opposite to an edge portion of the closed space.

5. The gene chip hybridization cassette according to any one of claims 1 to 4, characterized in that: The through hole is arranged to be opposite to the center position of the gene chip.

6. The gene chip hybridization cassette according to any one of claims 1 to 4, characterized in that: The predetermined interval is set so that when at least one gene chip is fixed on the substrate, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the cover facing the second surface is between 0.08 mm and 0.12 mm.

7. The gene chip hybridization cassette according to any one of claims 1 to 4, characterized in that: The gene chip hybridization box is further provided with at least one clamping device for releasably clamping the base plate, the spacer and the cover plate together.

8. The gene chip hybridization cassette according to any one of claims 1 to 4, characterized in that: A conductive element is provided in the gene chip placement region, and is provided so that when a first surface of the gene chip that does not include a probe region is bonded to the gene chip placement region, the conductive element is electrically connected to the first surface.

9. The gene chip hybridization cassette according to any one of claims 1 to 4, characterized in that: The substrate is formed of a conductive material so that when a first surface of a gene chip that does not include a probe region is bonded to the gene chip placement region, the conductive material is electrically connected to the first surface.

10. A gene chip hybridization box, characterized in that: The gene chip hybridization box comprises: substrate; A cover plate having at least one gene chip placement area, to which a first surface of the gene chip excluding the probe area can be bonded, the cover plate being provided with a through hole at a position outside and adjacent to the gene chip placement area, the gene chip placement area being provided with a recess for fixing and accommodating the gene chip in a tight fit; and a spacer disposed between the substrate and the cover plate, the spacer being used to create a predetermined gap between the substrate and the cover plate, and the spacer having an opening in a region facing the at least one gene chip arrangement region, thereby defining a gene chip accommodating space between the substrate and the cover plate; Wherein, the predetermined interval is set so that when the gene chip is fixed in the gene chip arrangement area, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the substrate facing the second surface is between 0.05mm and 0.15mm, so that when the hybridization liquid is injected through at least one through hole of the cover plate, the hybridization liquid can spread over the second surface of the gene chip by capillary action, and the cover plate is provided with a closed flange surrounding the gene chip arrangement area and the adjacent through holes. When the substrate, spacer and cover plate are fixed to each other, the closed flange abuts the substrate, thereby forming a closed space between the closed flange and the substrate; or the substrate is provided with a closed flange surrounding the gene chip arrangement area and the adjacent through holes. When the substrate, spacer and cover plate are fixed to each other, the closed flange abuts the cover plate, thereby forming a closed space between the closed flange and the cover plate.

11. The gene chip hybridization cassette according to claim 10, characterized in that: The cover plate is provided with at least two positioning hooks for fixing the gene chip in the gene chip arrangement area.

12. The gene chip hybridization cassette according to claim 10, characterized in that: The substrate and the spacer are formed integrally; or the cover and the spacer are formed integrally.

13. The gene chip hybridization cassette according to claim 10, characterized in that: The cover plate is provided with an exhaust hole communicating with the closed space.

14. The gene chip hybridization cassette according to claim 13, characterized in that: The through holes and the exhaust holes are arranged to be opposite to each other along a diagonal line of the gene chip arrangement area.

15. The gene chip hybridization cassette according to any one of claims 10 to 14, characterized in that: The predetermined interval is set so that when at least one gene chip is fixed on the substrate, the interval between the second surface of the gene chip opposite to the first surface and including the probe area and the surface of the cover facing the second surface is between 0.08 mm and 0.12 mm.

16. The gene chip hybridization cassette according to any one of claims 10 to 14, characterized in that: The gene chip hybridization box is further provided with at least one clamping device for releasably clamping the base plate, the spacer and the cover plate together.

17. The gene chip hybridization cassette according to any one of claims 10 to 14, characterized in that: A conductive element is provided in the gene chip placement region, and is provided so that when a first surface of the gene chip that does not include a probe region is bonded to the gene chip placement region, the conductive element is electrically connected to the first surface.

18. The gene chip hybridization cassette according to any one of claims 10 to 14, characterized in that: The cover plate is formed of a conductive material so that when a first surface of the gene chip that does not include a probe region is bonded to the gene chip placement area, the conductive material is electrically connected to the first surface.

19. A gene chip kit, characterized in that: The gene chip set includes: at least one gene chip; and The gene chip hybridization cassette according to any one of claims 1 to 18.

20. A gene chip hybridization method, characterized in that: The gene chip hybridization method comprises the following steps: Providing a gene chip hybridization cassette according to any one of claims 1 to 18; placing the gene chip to be hybridized in the gene chip hybridization box; and The hybridization solution is allowed to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and spread over the second surface of the gene chip containing the probe area through capillary action.

21. The gene chip hybridization method according to claim 20, characterized in that: The step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area by capillary action comprises: placing the gene chip hybridization box so that the substrate of the gene chip hybridization box is located below the cover plate; and The hybridization solution is injected into the gene chip hybridization box through the through hole of the gene chip hybridization box.

22. The gene chip hybridization method according to claim 20, characterized in that: The step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area by capillary action comprises: placing the gene chip hybridization box so that the cover of the gene chip hybridization box is located below the base plate; and The hybridization solution is injected into the gene chip hybridization box through the through hole of the gene chip hybridization box.

23. The gene chip hybridization method according to claim 20, characterized in that: The step of allowing the hybridization solution to enter the gene chip hybridization box through the through hole of the gene chip hybridization box and cover the second surface of the gene chip containing the probe area by capillary action comprises: placing the gene chip hybridization box so that the cover of the gene chip hybridization box is located below the base plate; and The cover plate is at least partially immersed in the hybridization solution, so that the hybridization solution is spread over the second surface of the gene chip containing the probe area through capillary action.

24. The gene chip hybridization method according to any one of claims 20 to 23, characterized in that: The method further comprises the following steps: An electric field is applied to the gene chip.

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