Patent Publication Number: US-2023151408-A1

Title: Gene sequencing structure, gene sequencing device and gene sequencing method

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application claims the priority of Chinese Patent Application No. 202111366541.2, filed on Nov. 18, 2021, the content of which is incorporated by reference in its entirety. 
     TECHNICAL FIELD 
     The present disclosure generally relates to the field of gene sequencing and bio-detection and, more particularly, relates to a gene sequencing structure, a gene sequence device, and a gene sequencing method. 
     BACKGROUND 
     Gene sequencing technology is a type of gene detection technology, which is the widely used in modern molecular biology research. Gene sequencing can analyze and determine the complete sequence of genes from blood or saliva to predict the possibility of suffering from a variety of diseases, such as cancer or leukemia. 
     Gene sequencing technologies mainly include the sequencing-by-synthesis technology, the ion-semiconductor sequencing technology, and the ligation sequencing technology, etc. Among them, the sequencing-by-synthesis technology is the mainstream, and occupies a leading position in the market. The sequencing-by-synthesis technology is to label four different deoxyribonucleotides with different colors of fluorescence (the four deoxyribonucleotides contain A adenine, T thymine, C cytosine, and G guanine). When a DNA polymerase synthesizes the complementary strand, each addition of a deoxyribonucleotide will release a different fluorescence. According to the captured fluorescence signal and processed by a specific a computer software, the sequence information of the to-be-tested DNA can be obtained. 
     At present, the existing fluorescence detection scheme requires a complicated laser light source and optical system, which complicates the sequencing system. In addition, labeling chemical reagents are extremely expensive, resulting in a high sequencing cost. Therefore, there is an urgent need for a gene sequencing to solve one or more problems set forth above and other problems in the arts. 
     SUMMARY 
     One aspect of the present disclosure provides a gene sequencing structure. The gene sequencing structure includes a substrate and a thin-film transistor array layer located on a side of the substrate. The thin-film transistor array layer includes a plurality of thin-film transistors, each of the plurality of thin-film transistors includes a first electrode, a second electrode, a third electrode, and a semiconductor layer, the first electrode and the second electrode are spaced apart and located in a same layer, the semiconductor layer is located on a side of the first electrode and the second electrode adjacent to the substrate, the semiconductor layer at least partially overlaps the first electrode and the second electrode, respectively, the semiconductor layer at least partially overlaps the third electrode, and an insulation layer is disposed between the semiconductor layer and the third electrode. The gene sequencing structure also includes an ion-sensitive layer located on a side of the semiconductor layer away from the substrate; and a micro-hole layer, located on a side of the ion-sensitive layer away from the substrate. The micro-hole layer includes a through-hole, the through-hole passes through the micro-hole layer in a direction perpendicular to the substrate, the through-hole at least partially overlaps the semiconductor layer, and the through-hole is used for receiving a to-be-tested single-stranded nucleic acid inside. Further, the gene sequencing structure includes a conductive structure located on a side of the layer away from the substrate, and electrically connected to the first electrode or the second electrode; and a detection chip, located on a side of the conductive structure away from the substrate, and electrically connected to the conductive structure. 
     Another aspect of the present disclosure provides a gene sequencing device. The gene sequencing device include a plurality of gene sequencing structures. Each gene sequencing structure includes a substrate and a thin-film transistor array layer located on one side of the substrate. The thin-film transistor array layer includes a plurality of thin-film transistors, each of the plurality of thin-film transistors includes a first electrode, a second electrode, a third electrode, and a semiconductor layer, the first electrode and the second electrode are spaced apart and located in a same layer, the semiconductor layer is located on a side of the first electrode and the second electrode adjacent to the substrate, the semiconductor layer at least partially overlaps the first electrode and the second electrode, respectively, the semiconductor layer at least partially overlaps the third electrode, and an insulation layer is disposed between the semiconductor layer and the third electrode. The gene sequencing structure also includes an ion-sensitive layer located on a side of the semiconductor layer away from the substrate; and a micro-hole layer, located on a side of the ion-sensitive layer away from the substrate. The micro-hole layer includes a through-hole, the through-hole passes through the micro-hole layer in a direction perpendicular to the substrate, the through-hole at least partially overlaps the semiconductor layer, and the through-hole is used for receiving a to-be-tested single-stranded nucleic acid inside. Further, the gene sequencing structure includes a conductive structure located on a side of the layer away from the substrate, and electrically connected to the first electrode or the second electrode; and a detection chip, located on a side of the conductive structure away from the substrate, and electrically connected to the conductive structure. 
     Another aspect of the present disclosure includes providing a gene sequencing method. The gene sequencing method includes providing a gene sequencing structure. The gene sequencing structure includes a substrate and a thin-film transistor array layer located on a side of the substrate. The thin-film transistor array layer includes a plurality of thin-film transistors, each of the plurality of thin-film transistors includes a first electrode, a second electrode, a third electrode, and a semiconductor layer, the first electrode and the second electrode are spaced apart and located in a same layer, the semiconductor layer is located on a side of the first electrode and the second electrode adjacent to the substrate, the semiconductor layer at least partially overlaps the first electrode and the second electrode, respectively, the semiconductor layer at least partially overlaps the third electrode, and an insulation layer is disposed between the semiconductor layer and the third electrode. The gene sequencing structure also includes an ion-sensitive layer located on a side of the semiconductor layer away from the substrate; and a micro-hole layer, located on a side of the ion-sensitive layer away from the substrate. The micro-hole layer includes a through-hole, the through-hole passes through the micro-hole layer in a direction perpendicular to the substrate, the through-hole at least partially overlaps the semiconductor layer, and the through-hole is used for placing a to-be-tested single-stranded nucleic acid inside. Further, the gene sequencing structure includes a conductive structure located on a side of the layer away from the substrate, and electrically connected to the first electrode or the second electrode; and a detection chip, located on a side of the conductive structure away from the substrate, and electrically connected to the conductive structure. The gene sequencing method also includes placing the to-be-tested single-stranded nucleic acid in the through-hole by contacting with the ion-sensitive layer; applying a first voltage signal to the first electrode or the second electrode, and applying a second voltage signal to the third electrode; passing four kinds of deoxyribonucleotides into the through-hole in sequence; detecting whether a current is generated between the first electrode and the second electrode through the conductive structure electrically connected to the detection chip using the detection chip to determine a base type of the to-be-tested single-stranded nucleic acid according to the deoxyribonucleotide introduced therein as the current is generated; and neutralizing and processing a reagent in the through-hole to restore the thin-film transistor to an initial state after each type of deoxyribonucleic acid is passed into the through-hole. The initial state is that the detection chip cannot detect a current generated between the first electrode and the second electrode. Further, the gene sequencing method includes eluting the reagent in the through-hole after the detection of each site of multiple to-be-tested single-stranded nucleic acids is completed. 
     Other aspects of the present disclosure can be understood by those skilled in the art in light of the description, the claims, and the drawings of the present disclosure. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The drawings incorporated in the specification and constituting a part of the specification illustrate the embodiments of the present disclosure, and together with the description are used to explain the principle of the present disclosure. 
         FIG.  1    illustrates a cross-sectional view of an exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure; 
         FIG.  2    illustrates a top view of an exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure; 
         FIG.  3    illustrates a cross-sectional view of another exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure; 
         FIG.  4    illustrates a cross-sectional view of another exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure; 
         FIG.  5    illustrates a cross-sectional view of another exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure; 
         FIG.  6    illustrates a cross-sectional view of another exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure; 
         FIG.  7    illustrates a cross-sectional view of an exemplary gene sequencing device according to various disclosed embodiments of the present disclosure; 
         FIG.  8    illustrates a top view of an exemplary gene sequencing device according to various disclosed embodiments of the present disclosure; and 
         FIG.  9    illustrates a flow chart of an exemplary gene sequencing method according to various disclosed embodiments of the present disclosure; and 
         FIG.  10    illustrates an exemplary variation diagram of a current between a first electrode and a second electrode according to various disclosed embodiments of the present disclosure. 
     
    
    
     DETAILED DESCRIPTION 
     To make the objectives, technical solutions, and advantages of the present disclosure clearer, the following further describes the present disclosure in detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present disclosure, and are not used to limit the present disclosure. 
     When certain words are used in the specification and claims to refer to specific components, those skilled in the art should understand that hardware manufacturers may use different terms to refer to the same component. This specification and claims do not use differences in names as a way of distinguishing components, but use differences in functions of components as a criterion for distinguishing. If “include” mentioned in the entire specification and claims is an open term, it should be interpreted as “include but not limited to”. “Approximately” means that within an acceptable error range, those skilled in the art can solve the technical problem within a certain error range, and basically achieve the technical effect. In addition, the term “coupled” herein includes any direct and indirect electrical coupling means. 
     Therefore, if it is described in the text that a first device is coupled to a second device, it may mean that the first device can be directly electrically coupled to the second device, or indirectly electrically coupled to the second device through other devices or coupling means. The following description of the specification is a preferred embodiment for implementing the application, but the description is for the purpose of explaining the general principles of the application and is not intended to limit the scope of the application. The scope of protection of this application shall be subject to those defined by the appended claims. Among them, the similarities between the embodiments will not be repeated one by one. 
     The present disclosure provides a gene sequencing structure, a gene sequencing device, and a gene sequencing method. 
       FIG.  1    illustrates a cross-sectional view of an exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure, and  FIG.  2    illustrates a top view of an exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure. 
     As shown in  FIGS.  1 - 2   , a gene sequencing structure  100  may include a substrate  10  and a thin-film transistor array layer  20 . The thin-film transistor array layer  20  may be located on one side of the substrate  10  and may include a plurality of thin-film transistors  70 . A thin-film transistor  70  may include a first electrode  71 , a second electrode  72 , a third electrode  73 , and a semiconductor layer  74 . The first electrode  71  and the second electrode  72  may be spaced apart and may be disposed in a same layer. The semiconductor layer  74  may be disposed on a side of the first electrode  71  and the second electrode  72  adjacent to the substrate  10 , and the semiconductor layer  74  may at least partially overlap the first electrode  71  and the second electrode  72 , respectively. The semiconductor layer  74  and the third electrode  73  may at least partially overlap, and an insulation layer  75  may be disposed between the semiconductor layer  74  and the third electrode  73 . 
     Further, the gene sequencing structure  100  may include an ion-sensitive layer  30 . The ion sensitive layer  30  may be located on a side of the semiconductor layer  74  away from the substrate  10 . 
     Further, the gene sequencing structure  100  may include a micro-hole layer  40 . The micro-hole layer  40  may be located on a side of the ion-sensitive layer  30  away from the substrate  10 . The micro-hole layer  40  may include a through-hole  41 . In the direction D 1  perpendicular to the substrate  10 , the through-hole  41  may pass through the micro-hole layer  40 , and the through-hole  41  may at least partially overlap the semiconductor layer  74 . The to-be-tested single-stranded nucleic acid  80  may be placed in the through-hole  41 . 
     Further, the gene sequencing structure  100  may include a conductive structure  50 . The conductive structure  50  may be located on a side of the micro-hole layer  40  away from the substrate  10 , and the conductive structure  50  may be electrically connected to the first electrode  71  or the second electrode  72 . 
     Further, the gene sequencing structure  100  may include a detection chip  60 . The detection chip  60  may be located on a side of the conductive structure  50  away from the substrate  10 , and may be electrically connected to the conductive structure  50 . 
     The gene sequencing structure in this embodiment may be configured to detect a gene sequence. For example, it may be configured to determine the types of bases of a set of to-be-tested single-stranded nucleic acids. Specifically, the conductive properties of the semiconductor layer in the thin-film transistor may be used to generate a current. Then, the unknown gene sequences may be determined through the detection of the detection chip. 
     Referring to  FIG.  1    and  FIG.  2   , the gene sequencing structure  100  may be entirely disposed on the substrate  10 . The substrate  10  may be configured as a carrier to carry other structures of the gene sequencing structure  100 . The substrate  10  may be a glass substrate, or a flexible substrate, which is not limited in this embodiment. The thin-film transistor array layer  20  disposed on one side of the substrate  10  may include the plurality of thin-film transistors  70 . 
     The thin-film transistor  70  may include the first electrode  71 , the second electrode  72 , the third electrode  73 , and the semiconductor layer  74 . In one embodiment, the first electrode  71  and the second electrode  72  may be source/drain metal layers, the third electrode  73  may be a gate metal layer, and the semiconductor layer  74  may be an active semiconductor layer. The first electrode  71  and the second electrode  72  may be arranged oppositely and spaced apart and located in the same layer. The semiconductor layer  74  may be located on the side of the first electrode  71  and the second electrode  72  adjacent to the substrate  10 . Along the direction D 1  perpendicular to the substrate  10 , the semiconductor layer  74  may at least partially overlap the first electrode  71  and the second electrode  72 , respectively. The insulation layer  75  may be disposed between the semiconductor layer  74  and the third electrode  73 , and along the direction D 1  perpendicular to the substrate  10 , the orthographic projections of the semiconductor layer  74  and the third electrode  73  on the substrate  10  may at least partially overlap. The thin-film transistor  70  provided in this embodiment may be configured to transmit a gene detection signal. 
     Further, an ion-sensitive layer  30  may be disposed on the side of the semiconductor layer  74  away from the substrate  10 . In one embodiment, the ion-sensitive layer  30  may include at least a sensitive material that can change the voltage of the ions generated by a base pairing. Accordingly, the ion-sensitive layer  30  in such an embodiment may need to be in contact with ions generated by the base pairing on the to-be-tested single-stranded nucleic acid  80 . In addition, the ion-sensitive layer  30  may also need to be in contact with the semiconductor layer  74 . The semiconductor layer  74  may be disposed in the ion-sensitive layer  30 . Accordingly, when the base pairing on the to-be-tested single-stranded nucleic acid  80  generates ions, the electric potential above the thin-film transistor  70  may be changed, and the conductivity of the semiconductor layer  74  may be changed. 
     Further, the micro-hole layer  40  may be disposed on the side of the ion-sensitive layer  30  away from the substrate  10 . The micro-hole layer  40  may include a plurality of through-holes  41 , and the through-holes  41  may pass through the micro-hole layer  40 , and the through-holes  41  may expose the ion-sensitive layer  30 . Along the direction D 1  perpendicular to the substrate  10 , the through-holes  41  and the semiconductor layer  74  may at least partially overlap. The through-holes  41  may be configured to host the to-be-tested single-stranded nucleic acid  80  to provide a certain amount of space for the base pairing of the to-be-tested single-stranded nucleic acid  80 . 
     It can be understood that, referring to  FIG.  2   , in one embodiment, a first voltage signal may be input to the first electrode  71  through the first metal wiring L 1 , and a second voltage signal may be input to the third electrode  73  through the second metal wiring L 2 . The signals may cause the surface of the semiconductor layer  74  to generate induced charges. As the first voltage signal input to the first electrode  71  changes, more induced charges may be generated on the surface of the semiconductor layer  74 . When the number of the induced charges generated on the surface of the semiconductor layer  74  reaches a certain level, a carrier transport channel may be formed between the first electrode  71  and the second electrode  72 . For example, a current may be generated between the first electrode  71  and the second electrode  72 . During the entire gene sequencing process, the thin-film transistor  70  may be operated in a linear region. Further, when different deoxyribonucleotides are sequentially injected into the through-holes  41  of the micro-hole layer  40  and paired with the bases of the to-be-tested single-strand nucleic acids  80 , hydrogen ions may be generated. The hydrogen ions may cause the potential above the thin-film transistor  70  to change, and the induced charges of the semiconductor layer  74  may be increased through the ion-sensitive layer  30 . Further, the current between the first electrode  71  and the second electrode  72  may be increased. By monitoring the change of the current between the first electrode  71  and the second electrode  72 , whether the base of the to-be-tested single-stranded nucleic acid  80  is paired may be determined, and the type of the base of the to-be-tested single-stranded nucleic acid  80  may also be determined. Comparing with the existing gene sequencing structure, the gene sequencing structure in this embodiment may adapt the existing thin-film transistor manufacturing process, and the base of the to-be-tested single-stranded nucleic acid may not require a fluorescent label and may not require a laser light source and optical system. In addition, the gene sequencing structure in this embodiment may add the signal control of the third electrode  73 , which may make the thin-film transistor  70  to be operated in the linear region such that the current change detection between the first electrode  71  and the second electrode  72  may be more obvious. 
     Further, referring to  FIG.  1    and  FIG.  2   , the conductive structure  50  may be disposed on the side of the micro-hole layer  40  away from the substrate  10 , and the detection chip  60  may be disposed on the side of the conductive structure  50  away from the substrate  10 . One end of the conductive structure  50  may be electrically connected to the second electrode  72 , and the other end of the conductive structure  50  may be electrically connected to the detection chip  60 . The detection chip  60  may detect the current change generated between the first electrode  71  and the second electrode  72  through the conductive structure  50 . On the one hand, by introducing the conductive structure  50 , the detection positions of the thin-film transistor  70  may be effectively increased, and the experimental efficiency may be effectively improved. On the other hand, the detection chip  60  may be disposed on the side of the conductive structure  50  away from the substrate  10 , which may effectively save space of the side of the conductive structure  50 . 
     It should be noted that the embodiment shown in  FIG.  1    only schematically shows a schematic diagram of the conductive structure  50  being electrically connected to the thin-film transistor  70  and the detection chip  60 , respectively, and may not represent the actual size of the conductive structure  50 . Further,  FIG.  1    only shows a form in which two adjacent gene detection structures are arranged side by side. Further, the embodiment shown in  FIG.  1    only schematically shows one shape of the conductive structure  50 , the conductive structure  50  may also have other shapes, which is not limited in this disclosure. The embodiment shown in  FIG.  2    only schematically shows the positional relationship between the first electrode  71 , the second electrode  72 , the third electrode  73 , the semiconductor layer  74 , and the conductive structure  50 , and may not represent the actual sizes. 
     Further, referring to  FIG.  1   , in one embodiment of the present disclosure, the gene sequencing structure may further include an output detection terminal  90 . The output detection terminal  90  may be disposed between the conductive structure  50  and the first electrode  71  or the second electrode  72 . The conductive structure  50  may be electrically connected with the first electrode  71  or the second electrode  72  through the output detection terminal  90 . 
     For example, referring to  FIGS.  1 - 2   , the gene sequencing structure  100  in this embodiment may include the output detection terminal  90 . The output detection terminal  90  may be disposed between the conductive structure  50  and the first electrode  71  or the second electrode  72 . The output detection terminal  90  may match the size and shape of the conductive structure  50  without affecting the characteristics of the thin-film transistor  70 . Further, the output detection terminal  90  may be selected to have a material different from the first electrode  71  or the second electrode  71  of the thin-film transistor  70 . For example, the output detection terminal  90  may be made of a material with a better corrosion resistance to protect the thin-film transistor  70 . In one embodiment, when the first electrode  71  is configured to input the first voltage signal, the output detection terminal  90  may be electrically connected to the second electrode  72  and the conductive structure  50 , respectively. When the second electrode  72  is configured to input the first voltage signal, the output detection terminal  90  may be electrically connected to the first electrode  71  and the conductive structure  50 , respectively. As shown in  FIG.  1   , the output detection terminal  90  may be electrically connected to the second electrode  72 ; and the first voltage signal may input by the first electrode  71  through the first metal wiring L 1 . In such a way, the detection chip  60  may detect the current change between the first electrode  71  and the second electrode  72  through the conductive structure  50  and the output detection terminal  90 , which may effectively increase the detection positions of the thin-film transistor  70  and improve the detection efficiency. 
     It should be noted that the embodiment shown in  FIG.  1    only schematically shows that the cross-section of the output detection terminal  90  is an L-shaped structure, and may not represent the actual size of the output detection terminal  90 . The structure of the output detection terminal  90  may also be other structures that are not limited in this disclosure. 
       FIG.  3    illustrates a cross-sectional view of another exemplary gene sequencing structure consistent with various disclosed embodiments of the present disclosure. As shown in  FIG.  3    and referring to  FIG.  1   , in one embodiment of the present disclosure, the output detection terminal  90  may include a first branch  91  and a second branch  92 . The first branch  91  may be located on the side of the micro-hole layer  40  away from the substrate  10 , and the first branch  91  may be in contact with the conductive structure  50 . The second branch  92  may be located in the micro-hole layer  40 , and a portion of the second branch  92  may be located in the ion-sensitive layer  30  and may be in contact with the first electrode  71  or the second electrode  72 . 
     Referring to  FIG.  1    and  FIG.  3   , in one embodiment, the output detection terminal  90  may include two portions: the first branch  91  and the second branch  92 . The first branch  91  and the second branch  92  may be a same structure and may be formed at the same time. For example, a contact through-hole may be formed in the micro-hole layer  40  and the ion-sensitive layer  30 , the second branch  92  may be formed in the contact through-hole, and the second branch  92  may be electrically in contact with the first electrode  71  or the second electrode  72 . The first branch  91  may be located on the side of the micro-hole layer  40  away from the substrate  10 , and the first branch  91  may be electrically connected to the conductive structure  50 . In such an embodiment, the second branch  92  may be electrically connected with the second electrode  72 . In another embodiment, as shown in  FIG.  1   , a portion of the first branch  91  may be formed in the through-hole  41 , and a contact through-hole may not need to be formed in the micro-hole layer  40 , and the fabrication process may be simplified. In some embodiments, as shown in  FIG.  3   , the contact through-hole may pass through the ion-sensitive layer  30 . Under such a configuration, the contact through-hole may need to be formed in the ion-sensitive layer  30 . In this embodiment, through the first branch  91  and the second branch  92  of the detection terminal  90 , the conductive structure  50  and the detection chip  60  may be arranged on the side of the thin-film transistor array layer  20  away from the substrate  10 . On the one hand, it may effectively save the space on the side of the substrate  10  away from the array layer  20  of the thin-film transistor  70 . On the other hand, it may also effectively reduce the manufacturing cost of the detection chip  60 . 
       FIG.  4    illustrates a cross-sectional view of another exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure. As shown in  FIG.  4   , in one embodiment of the present disclosure, the gene sequencing structure  100  may further includes a counter substrate  110  located on the side of the conductive structure  50  away from the substrate  10 . 
     The detection chip  60  may be located on the side of the counter substrate  110  away from the conductive structure  50 . The counter substrate  110  may include a plurality of conductive hole structures  111  which may be conductive, and the conductive hole structures  111  may be electrically connected to the conductive structure  50 . The detection chip  60  may be electrically connected to the conductive hole structures  111  through the first signal wirings  112 . 
     Referring to  FIG.  4   , the gene sequencing structure  100  in this embodiment may also include the counter substrate  110 . In one embodiment, the counter substrate  110  may be a glass substrate, a PCB substrate, or an encapsulation layer, etc. The counter substrate  110  may be located on the side of the conductive structure  50  away from the substrate  10 . The counter substrate  110  may be disposed opposite to the substrate  10 . The thin-film transistor array layer  20 , the ion-sensitive layer  30 , the micro-hole layer  40 , and the conductive structure  50  may all be located between the counter substrate  110  and the substrate  10 . In one embodiment, the detection chip  60  may be located on the side of the counter substrate  110  away from the conductive structure  50 , and the plurality of conductive hole structures  111  may be formed in the counter substrate  110 . For example, holes may be formed in the counter substrate  110 , and a conductive material may be injected into the holes to electrically connect the detection chip  60  and the conductive structure  50 . In one embodiment, the conductive material injected into the holes may be made of a same material as the conductive structure  50 . 
     The side of the conductive hole structures  111  adjacent to the substrate  10  may be electrically connected to the conductive structure  50 , and the side of the conductive hole structures  111  away from the substrate  10  may be electrically connected to the detection chip  60  through the first signal wiring  112 . In such a manner, the detection chip  60  and the conductive structure  50  may be electrically connected; and the detection chip  60  may detect the current change between the first electrode  71  and the second electrode  72 . Further, the counter substrate  110  may be in the form of a jig and integrated in the external machine; and may not be used as a consumable, the production cost may be effectively reduced. 
     It should be noted that the embodiment shown in  FIG.  4    only schematically shows the structural diagram of the conductive hole structures  111 , and may not represent the actual size of the conductive hole structures  111 . 
     Further, referring to  FIG.  4   , in another embodiment of the present disclosure, the gene sequencing structure may further include an encapsulation structure  120 . The encapsulation structure  120  may be located on the side surface of the stacked structure having the substrate  10 , the thin-film transistor array layer  20 , the ion-sensitive layer  30 , and the micro-hole layer  40 , the conductive structure  50  and the opposite substrate  110 . 
     Referring to  FIG.  4   , the gene sequencing structure  100  in this embodiment may also include the encapsulation structure  120 . The encapsulation structure  120  may encapsulate the stacked structure having the substrate  10 , the thin-film transistor array layer  20 , the ion-sensitive layer  30 , the micro-hole layer  40 , the conductive structure  50  and the counter substrate  110 . In one embodiment, the encapsulation structure  120  may be located on the side surface of the above stacked structure. Along the direction D 2  parallel to the substrate  10 , the detection chip  60  may be bound to the side of the counter substrate  110  away from the substrate  10 . Such a configure may effectively save the space on the side of the substrate  10  away from the counter substrate  110 . 
     It should be noted that the embodiment shown in  FIG.  4    only schematically shows a schematic diagram of the encapsulation structure  120  encapsulating two gene sequencing structures. In some embodiments, the encapsulation structure  120  may also encapsulate multiple gene sequencing structures, which is not limited in this disclosure. 
       FIG.  5    illustrates a cross-sectional view of another exemplary gene sequencing structure accordingly to various disclosed embodiments of the present disclosure, and  FIG.  6    shows a schematic top view of another exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure. As shown in  FIG.  5    and  FIG.  6   , in one embodiment of the present disclosure, the gene sequencing structure  100  may also include a counter substrate  110 . The counter substrate  110  may be located on the side of the conductive structure  50  away from the substrate  10 . 
     The detection chip  60  may be located on the side of the counter substrate  110  adjacent to the conductive structure  50 . The detection chip  60  and the conductive structure  50  may be electrically connected through the second signal wiring  113 . 
     Referring to  FIGS.  5 - 6   , the gene sequencing structure  100  in this embodiment may include the counter substrate  110 . The counter substrate  110  may be a glass substrate  10 , a PCB substrate  10 , or an encapsulation structure, etc. The counter substrate  110  may be located on the side of the conductive structure  50  away from the substrate  10 . In one embodiment, the conductive structure  50  may be fabricated in the counter substrate  110 . The counter substrate  110  may be arranged opposite to the substrate  10 . The thin-film transistor array layer  20 , the ion-sensitive layer  30 , the micro-hole layer  40 , the conductive structure  50 , and the detection chip  60  may all be disposed between the counter substrate  110  and the substrate  10 . The detection chip  60  may be electrically connected to the conductive structure  50  through the second signal wiring  113  to achieve the electrical connection between the detection chip  60  and the conductive structure  50 . The detection chip  60  may detect the current change between the first electrode  71  and the second electrode  72 . Further, the counter substrate  110  may be in the form of a jig, may be integrated in an external machine, and may not be used as a consumable. Accordingly, the cost may be reduced. 
       FIG.  7    illustrates a cross-sectional view of anther exemplary gene sequencing structure according to various disclosed embodiments of the present disclosure. As shown in 
       FIG.  7   , in one embodiment of the present disclosure, a first insulation layer  114  may be disposed among a plurality of second signal wirings  113 . 
     Referring to  FIG.  7   , in this embodiment, in view of the large number of second signal wirings  113 , the plurality of second signal wirings  113  may be disposed in different layers. The first insulation layer  114  may be disposed between the detection chip  60  and the second signal wirings  113  of the conductive structure  50 , and the second signal wirings  113  in different layers may be electrically connected through vias. In such a configuration, the electrical connection between detection chip  60  and the conductive structure  50  may be achieved, while the concentrated wiring of the second signal wirings  113  may be avoided to avoid a poor load. 
     It should be noted that the embodiment shown in  FIG.  7    only schematically shows a structure diagram provided with two layers of second signal wirings  113 , multiple layers of second signal wirings  113  may also be provided, and an insulation layer may be disposed among the multiple layers of second signal wirings  113 , which is not limited in the present disclosure. 
     Further referring to  FIG.  5    and  FIG.  7   , in anther embodiment of the present disclosure, the gene sequencing structure  100  may further include the encapsulation structure  120 . The encapsulation structure  120  may be disposed on the side surfaces of a stacked structure having the substrate  10 , the thin-film transistor array layer  20 , the ion-sensitive layer  30 , the micro-hole layer  40 , the conductive structure  50 , the detection chip  60  and the counter substrate  110 . 
     Referring to  FIG.  5    and  FIG.  7   , in the gene sequencing structure  100  in this embodiment, the encapsulation structure  120  may encapsulate the stacked structure including the substrate  10 , the thin-film transistor array layer  20 , the ion-sensitive layer  30 , the micro-hole layer  40 , the conductive structure  50 , the detection chip  60  and the counter substrate  110 . In one embodiment, the encapsulation structure  120  may be located on the side surface of the stacked structure and the side surface D 2  parallel to the substrate  100 . In such a manner, the detection chip  60  may be bound to the side of the counter substrate  110  away from the substrate  10 . Accordingly, the space on the side of the substrate  10  away from the counter substrate  110  may be effectively saved. 
     Further, referring to  FIG.  4    to  FIG.  7   , in one embodiment of the present disclosure, the encapsulation structure  120  may include a plurality of openings  130 . 
     Referring to  FIGS.  4 - 7   , the encapsulation structure  120  in this embodiment may include a plurality of openings  130 . The openings  130  may be located at any position of the encapsulation structure  120 . The openings  130  may be used to inject a single-stranded nucleic acid into the through-hole  41  of the micro-hole layer  40 , and it may also be used to inject pairing bases into the through-hole  41 , and it may also discharge the unnecessary solution in the through-hole  41 . For example, the opening  130  may be used for injecting or discharging a solution into the through-hole  41  of the micro-hole layer  40 . 
     It should be noted that the embodiments shown in  FIGS.  4  to  7    only schematically show one position where the opening is located. The opening may also be located at other positions, which is not limited in the present disclosure. 
     Further, referring to  FIG.  1   , in embodiment of the present disclosure, the diameter of the through-hole  41  may be in a range of approximately 3 μm - 6 μm. 
     Referring to  FIG.  1   , the diameter of the through-hole  41  in this embodiment may be in a range of approximately 3 μm-6 μm. In one embodiment, the diameter of the through-hole  41  may also be 4 μm or 5 μm. If the diameter of the through-hole  41  is greater than 6 μm, it may affect the number of through-holes  41 . If the diameter of the through-hole  41  is smaller than 3 μm, it may be difficult to achieve by using the panel technology. By limiting the diameter of the through-holes  41 , there may be more detection positions integrated in the gene sequencing structure  100 . Accordingly, the detection efficiency may be improved. 
     It should be noted that the embodiment shown in  FIG.  1    only schematically shows the structural diagram of the through-holes  41 , and may not represent the actual size of the through-holes  41 . 
     Further, referring to  FIG.  1   , in another embodiment of the present disclosure, along the direction D 1  perpendicular to the substrate  10 , the height of the conductive structure  50  may be in a range of approximately 3 μm-5 μm. 
     Referring to  FIG.  1   , in such an embodiment, along the direction D 1  perpendicular to the substrate  100 , the height of the conductive structure  50  may be in a range of approximately 3 μm-5 μm. In one embodiment, the height of the conductive structure  50  may be approximately 4 μm. In such a case, the substrate  10  may be a glass substrate. In such a configuration, the patterning accuracy of the conductive structure  50  may be higher, which may facilitate the realization of the requirement for the detection positions of the high-density thin-film transistor  70   s  in the present disclosure. 
     It should be noted that the embodiment shown in  FIG.  1    only schematically shows a schematic structural diagram of the conductive structure  50  and may not represent the actual size of the conductive structure  50 . 
     In one embodiment of the present application, the material of the conductive structure  50  may be electroplated copper. 
     For example, the conductive structure  50  in this embodiment may be made of electroplated copper. When the conductive structure  50  is electroplated with a height of 3 μm to 5 μm in the direction D 1  perpendicular to the substrate  10 , a highly accurate conductive structure  50  may be realized. At this time, the substrate  10  may be a glass substrate. When the conductive structure  50  is electroplated with copper with a height of 5 μm to 50 μm in the direction D 1  perpendicular to the substrate  100 , the requirements for the production accuracy may be relatively low. At this time, the substrate  10  may be a PCB substrate, the warpage of the substrate  10  may be avoided. 
     Further, referring to  FIG.  1   , in another embodiment of the present disclosure, the thin-film transistor  70  may be a bottom-gated structure. 
     Referring to  FIG.  1   , the thin-film transistor  70  in this embodiment may be a bottom-gated structure. It may be understandable that the third electrode  73  in this embodiment may be the gate of the thin-film transistor  70 . For example, the gate may be located on the semiconductor layer  74  adjacent to the substrate  10 . In such a configuration, the potential change generated above the thin-film transistor  70  may not be shielded by the third electrode  73 , i.e., the gate, but the conductivity of the semiconductor layer  74  may be changed by the ion-sensitive layer  30 . 
     In one embodiment of the present disclosure, the material of the ion-sensitive layer  30  layer may be silicon nitride, or silicon oxide, etc. 
     The material of the ion-sensitive layer  30  in this embodiment may be silicon nitride or silicon oxide. The ion-sensitive layer  30  may be used to sense changes in ions and to sense the occurrence of the base pairing. The ion-sensitive layer  30  may be formed by hydrogen-ion-sensitive material, or the ion-sensitive material a voltage of which may be changed by the ions generated in the base pairing. Silicon nitride or silicon oxide may achieve the required functions of the ion-sensitive layer  30 . 
     The present disclosure also provides a gene sequencing device.  FIG.  8    illustrates a cross-sectional view of an exemplary gene sequencing device according to various disclosed embodiments of the present disclosure, and  FIG.  9    illustrates a top view of an exemplary gene sequencing device according to various disclosed embodiments of the present disclosure. 
     As shown in  FIG.  8    and  FIG.  9   , the gene sequencing device  200  may include a plurality of gene sequencing structures  100 . The gene sequencing structure  100  may be the gene sequencing structure provided by any embodiment of the present disclosure. In the gene sequencing device  200  of the present disclosure, a plurality of gene sequencing structures  100  may be arranged in an array. 
     Referring to  FIG.  8    and  FIG.  9   , the plurality of gene sequencing structures  100  may be arranged as an array or may be arranged uniformly. They may also be arranged in other ways, which are not limited in this application. In different gene sequencing structures  200 , all the first electrodes  71  or the second electrodes  72  may input a same first voltage signal through the first metal wiring L 1 , and all the third electrodes  73  may input a same second voltage signal through the second metal wiring L 2 . The connection lines of the first electrodes  71  or the second electrodes  72  in the gene sequencing structures  100  located in a same row or a same column may be connected in parallel. The connection lines of the third electrodes  73  in the gene sequencing structures  100  located in a same row or a same column may be connected in parallel. In this embodiment, the first electrodes  71  of the gene sequencing structures  100  located in the same row may be connected in parallel, and the third electrodes  73  of the gene sequencing structures  100  located the same row may be connected in parallel. On the one hand, the gene sequencing structures  100  arranged in the array may be able to improve the efficiency of the gene sequencing. On the other hand, the gene sequencing structures  100  may be used to sense whether the bases on the to-be-tested nucleic acid single strand  80  are paired, the gene sequencing may be simple, and the cost of the gene sequencing may be effectively reduced. 
     Further, the present disclosure provides a gene sequencing method.  FIG.  10    illustrates a flow chart of an exemplary gene sequencing method according to various disclosed embodiment of the present disclosure.  FIG.  11    is a variation curve of the current between the electrode and the second electrode according to various disclosed embodiments of the present disclosure. The gene sequencing method may use the present disclosed gene sequence structure and gene sequencing device to perform a gene sequencing. As shown in  FIGS.  10 - 11   , and referring to  FIG.  1   , the gene sequencing method may include:
         in S 101 : placing a to-be-tested single-stranded nucleic acid  80  in the through-hole  41  by contacting with the ion-sensitive layer  30 ;   in S 102 : applying a first voltage signal to the first electrode  71  or the second electrode  72 , and applying a second voltage signal to the third electrode  73 ;   in S 103 : passing four kinds of deoxyribonucleotides into the through-hole  41  in sequence;   in S 104 : detecting whether a current is generated between the first electrode  71  and the second electrode  72  through the conductive structure  50  electrically connected to the detection chip  60  using the detection chip  60  to determine the base type of the to-be-tested single-stranded nucleic acid  80  according to the deoxyribonucleotide introduced therein as the current is generated;   in S 105 : neutralizing and processing the reagent in the through-hole  41  to restore the thin-film transistor  70  to the initial state after each type of deoxyribonucleic acid is passed into the through-hole  41 , wherein the initial state is that the detection chip  60  cannot detect the current generated between the first electrode  71  and the second electrode  72 ; and   in S 106 : eluting the reagent in the through-hole  41  after the detection of each site of multiple to-be-tested single-stranded nucleic acids  80  is completed.       

     Referring to  FIG.  10    and  FIG.  11    and referring to  FIG.  1    and  FIG.  2   , the present disclosed gene sequencing method may utilize the present disclosed gene sequencing structure. Thus, the gene sequencing method may be relatively simple, and the cost of gene sequencing method may be reduced. 
     In S 101  of the gene sequencing method, different to-be-tested single-stranded nucleic acids  80  may be loaded into different through-holes  41  through the openings  130  on the sidewalls of the encapsulation structure  120 , and the to-be-tested single-stranded nucleic acids  80  may be in touch with the ion-sensitive layer  30  exposed by the through-holes  41 . 
     In S 102 , a first voltage signal may be applied to the first electrode  71 , or may be applied to the second electrode  72 , and a second voltage signal may be applied to the third electrode  73 . The first voltage signal and the second voltage signal may be same or different. In one embodiment, the first voltage signal may be applied to the first electrode  71  through the first metal wiring L 1 , and the second voltage signal may be applied to the third electrode  73  through the second metal wiring L 2 . 
     In S 103 , an adenine may be injected into different through-holes  41  through the openings  130  on the sidewalls of the encapsulation structure  120 . If the first site of the single-stranded nucleic acid  80  in the through-hole  41  is paired, hydrogen ions may be released, which may cause the potential on the semiconductor layer  74  to change, and whether the current between the first electrode  71  and the second electrode  72  changes may be detected by the detection chip  60 . If the current changes, the pairing is successful, and it may be determined that the base pairing is successful. The type of the base of the first site of the single-stranded nucleic acid  80  in the through-hole  41  may be determined as T thymine. Then, the T thymine may be injected into the through-hole  41  to further determine the base of the single-stranded nucleic acid  80  in the through-hole  41  to the A adenine. Sequentially, the C cytosine and G guanine may be injected into the through-holes  41  until the bases at the first sites of the single-stranded nucleic acids  80  in all the through-holes  41  match. At this time, the reagent in the through-hole  41  may be eluted to prepare for the gene detection of the single-stranded nucleic acid  80  at the second site in the through-hole  41 . The specific detection method may be same as that of the first site. 
     In S 104 , in the above steps, when the deoxyribonucleotides passing through the through-hole  41  successfully pairs with the bases of the to-be-tested single-stranded nucleic acids  80 , the detection chip  60  may detect whether a current generated between the first electrode  71  and the first electrode  71  through the conductive structure  50  changes. If the current generated between the first electrode  71  and the second electrode  72  changes, the base pairing may be successful. The type of the base of the to-be-tested single-stranded nucleic acid  80  may be determined according to the type of the deoxyribonucleotide passed in. 
     In S 105 , in the above steps, after each type of deoxyribonucleotide is passed into the through-hole  41 , the reagent in the through-hole  41  may need to be neutralized to restore the thin-film transistor  70  to the initial state. The initial state of the thin-film transistor  70  may be that the detection chip  60  may not detect the current generated between the first electrode  71  and the second electrode  72 . When the bases of the to-be-tested single-stranded nucleic acid  80  are paired, the concentration of hydrogen ions in the through-hole  41  may change. If it is not processed in time, it may affect the concentration of hydrogen ions generated after the next deoxyribonucleotide passes into the through-hole  41  and pairs with the with the to-be-tested single-stranded nucleic acid  80  and may further affect the current change between the first electrode  71  and the second electrode  72 . Accordingly, the accuracy of this pairing detection may be affected. Therefore, after each pairing happens on the base of the to-be-tested single-stranded nucleic acid  80 , the reagent in the through-hole  41  may need to be neutralized to avoid causing the subsequent errors. 
     In S 106 , in the above steps, the single-stranded nucleic acid  80  in the through-hole  41  includes a plurality of site bases. After the detection of the first sites of the single-stranded nucleic acids  80  in all the through-holes  41  is completed, the solution in the through-holes  41  may need to be eluted to ensure that the reagent in the through-holes  41  may return to the original state, and may not affect the detection of the second sites of the single stranded nucleic acids  80  in all the through-holes  41 . 
     In one embodiment, the first voltage signal may be applied to the first electrode  71  through the first metal wiring L 1 , and the second voltage signal may be applied to the second electrode  72  through the second metal wiring L 2 . A certain current may be generated between the first electrode  71  and the second electrode  72  and the thin-film transistor  70  may be operated in the linear region. On this basis, after the bases of the to-be-tested single-stranded nucleic acids  80  in the through-holes  41  are paired, a potential change may be generated above the thin-film transistor  70 . The conductivity of the semiconductor layer  74  may be further changed to make the current change between the first electrode  71  and the second electrode  72  more obvious. 
     Referring to the current change between the first electrode  71  and the second electrode  72  shown in  FIG.  11   , the thick solid line illustrates the successful base pairing on the tested single-stranded nucleic acid  80 , and the thin solid line illustrates the unsuccessful base pairing on the tested single-stranded nucleic acid  80 . The abscissa in  FIG.  11    is the voltage of the semiconductor layer  74 , the ordinate is the current between the first electrode  71  and the second electrode  72 . Through the above sequencing method, the detection of the gene sequence may be more convenient and efficient. Further, in this embodiment, after all the bases of the to-be-tested single-stranded nucleic acids  80  are tested, the reagent in the through-hole  41  may be eluted, which may ensure the accuracy of the gene sequencing at the next site of the single-stranded nucleic acid  80 . 
     In one embodiment of the present disclosure, neutralizing the reagent in the through-holes  41  may include following steps. 
     After the base of the to-be-tested single-stranded nucleic acid  80  in the through-hole  41  is paired with the deoxyribonucleotides introduced into the through-hole  41 , a phosphate buffer solution may be injected into the through-hole  41  through the opening  130  to neutralize the hydrogen ions in the through-hole  41  to restore the hydrogen ion concentration index in the through-hole  41  to the value before the introduction of deoxyribonucleotides. 
     In one embodiment, the phosphate buffer solution may be used to neutralize the reagent in the through-hole  41  such that the concentration index of hydrogen ions in the through-hole  41  may not affect the continuation of the pairing of the bases on the to-be-tested single-stranded nucleic acid, and the accuracy of gene sequencing results may be ensured. 
     It can be seen from the above embodiments that the present disclosed gene sequencing structure, gene sequencing device and gene sequencing method may have the following beneficial effects. 
     The gene sequencing structure, gene sequencing device and gene sequencing method provided by the present disclosure may be provided with a substrate, a thin-film transistor array layer, an ion-sensitive layer, a micro-hole layer, a conductive structure, and a detection chip. The successful base pairing may cause a change in the potential above the thin-film transistor. The conductivity of the semiconductor layer in the thin-film transistor may be used to generate a current between the first electrode and the second electrode. The change in the current may be detected by the detection chip to detect the sequence of the gene. The base of the to-be-detected single-stranded nucleic acid may not require a fluorescent labeling, laser light source and optical system. Further, the signal control of the third electrode is added in this disclosure to make the thin-film transistor being operated in the linear region. Accordingly, the current change detection between the first electrode and the second electrode may be more obvious. Further, the present disclosure may also introduce a conductive structure, which may effectively increase the detection positions of the thin-film transistor, and may effectively improve the experimental efficiency. On the other hand, the detection chip may be arranged on the side of the conductive structure away from the substrate, which may effectively save the space on the side of the substrate. 
     The above are only specific embodiments of the present disclosure, but the protection scope of the present disclosure is not limited thereto. Any person skilled in the art can easily think of various equivalent modifications or changes within the technical scope disclosed in the present disclosure. Equivalent modifications or replacements should all be covered within the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure should be subject to the protection scope of the claims.