Gene sequencing system, gene sequencing method and sample vector

Abstract

The invention belongs to the technical field of gene sequencing, and provides a gene sequencing system, a gene sequencing method and a sample carrier in order to solve the problem of limited efficiency and flux in the prior art. The sample carrier has two basically parallel sample attachment surfaces, a swimming lane is formed between the two attachment surfaces, the swimming lane allows fluid to pass through or stay, the three-dimensional space can be fully utilized, more samples can be borne, the utilization rate of the sample carrier can be improved, the limitation of mutual interference of fluorescence signals can be broken through, and the sequencing flux can be improved. The gene sequencing system comprises the sample carrier, a fluid module, a collection module and an analysis module, provides an operation platform for the double-layer sample carrier, can realize a gene sequencing function, and has the advantages of high efficiency and large flux. The gene sequencing method comprises the steps of loading a sequencing sample, providing a sequencing reagent, collecting a fluorescence signal and outputting sequence information, provides method guidance for application of the double-layer sample carrier, and can efficiently complete sequencing operation.

Description

Technical Field

[0001] The present application relates to the field of gene sequencing technology, and in particular to a gene sequencing system, a gene sequencing method and a sample carrier. Background Art

[0002] DNA sequencing, also known as DNA sequencing or gene sequencing, refers to the analysis of the base sequence of a specific DNA fragment, specifically the arrangement of adenine (A), thymine (T), cytosine (C), and guanine (G). Currently, the most commonly used sequencing method involves attaching the sample to be sequenced (such as a sequencing template) to a carrier. A fluorescently labeled probe (such as dNTP) is then delivered to the sample. The sample binds to the probe, and the fluorescent marker is excited and emits light. This "binding" and "luminescence" process continues in a continuous cycle. By collecting and analyzing the fluorescent signal emitted after each binding, the base sequence information of the sample to be sequenced is ultimately determined.

[0003] Existing technologies typically attach all samples to a single surface. If the samples are too close together, fluorescence signals can interfere with each other, affecting the final sequencing results. Therefore, the sample layout density cannot be too high. However, this layout makes it difficult to further improve the utilization of sample carriers, limiting sequencing efficiency and throughput. "Throughput" generally refers to the amount of data generated per unit time or in a single sequencing run. Summary of the Invention

[0004] The purpose of this application is to overcome the shortcomings of the existing technology and provide a gene sequencing system, a gene sequencing method and a sample carrier, which have high sequencing efficiency and high throughput.

[0005] A gene sequencing system provided in the present application includes a sample carrier, a fluid module, a collection module and an analysis module. The sample carrier is used to carry sequencing samples. The fluid module can provide the samples with reagents required for sequencing reactions. The collection module is used to collect fluorescent signals emitted by the samples. The analysis module is used to analyze the fluorescent signals collected by the collection module and output sequence information. The sample carrier includes a first substrate and a second substrate. The first substrate has a first attachment surface, and the second substrate has a second attachment surface. The first attachment surface and the second attachment surface are arranged opposite to each other and are basically parallel. A swimming lane is formed between the first attachment surface and the second attachment surface. The swimming lane allows fluid to pass through or stay. Multiple samples are respectively attached to the first attachment surface and the second attachment surface.

[0006] The gene sequencing system provided in this application uses a sample carrier with a double-layer sample attachment surface, which has the advantage of high throughput. At the same time, the system includes a fluid module, a collection module and an analysis module, which cooperates with the double-layer sample carrier to automatically complete the sequencing work, and has the characteristics of high efficiency.

[0007] Furthermore, as an implementation method of the relative position relationship of samples on the two attachment surfaces of the sample carrier, the vertical projection of the sample on the first attachment surface on the second attachment surface will not overlap with the sample on the second attachment surface, or the vertical projection of the sample on the second attachment surface on the first attachment surface will not overlap with the sample on the first attachment surface.

[0008] Furthermore, as another embodiment of the relative position relationship of samples on the two attachment surfaces of the sample carrier, the vertical projection of the sample on the first attachment surface on the second attachment surface overlaps with the sample on the second attachment surface, or the vertical projection of the sample on the second attachment surface on the first attachment surface overlaps with the sample on the first attachment surface.

[0009] Furthermore, as an arrangement of samples on a single attachment surface of the sample carrier, the samples on the first attachment surface and / or the second attachment surface are arranged in a matrix.

[0010] Furthermore, as another arrangement of samples on a single attachment surface of the sample carrier, the samples on the first attachment surface and / or the second attachment surface are arranged in a staggered manner.

[0011] Furthermore, as a third arrangement of samples on a single attachment surface of the sample carrier, the samples on the first attachment surface and / or the second attachment surface are randomly distributed.

[0012] Furthermore, the first attachment surface and the second attachment surface have attachment sites for placing samples, and the attachment sites on the first attachment surface and / or the second attachment surface are arranged in a matrix.

[0013] Furthermore, the first attachment surface and the second attachment surface have attachment sites for placing samples, and the attachment sites on the first attachment surface and / or the second attachment surface are arranged in a staggered manner.

[0014] Furthermore, the first attachment surface and the second attachment surface have attachment sites for placing samples, and the attachment sites on the first attachment surface and / or the second attachment surface are randomly distributed.

[0015] Furthermore, the collection module collects the fluorescence signal on one side of the sample carrier.

[0016] Furthermore, the collection module collects the fluorescence signal on both sides of the sample carrier.

[0017] Furthermore, the acquisition module includes multiple acquisition sub-modules.

[0018] Furthermore, the acquisition module uses super-resolution imaging technology to acquire the fluorescence signal.

[0019] Furthermore, the acquisition module includes a CCD camera or a CMOS camera, or the acquisition module includes an area array camera, a line array camera or a TDI camera.

[0020] Furthermore, the gene sequencing system includes a plurality of the sample carriers.

[0021] Furthermore, the sample carrier includes a plurality of sample sub-carriers.

[0022] Furthermore, in order to achieve a more complete sequencing function, the gene sequencing system also includes a sample preparation module, which is used to prepare samples that can be loaded into the sample carrier for sequencing.

[0023] Furthermore, the sample preparation module includes a plurality of sample preparation submodules.

[0024] Furthermore, the sample prepared by the sample preparation module is transferred to the sample carrier through a mechanical automation device or the fluid module.

[0025] Furthermore, the sample carrier is associated with the fluid module in a contact-type or non-contact-type manner, the acquisition module is associated with the sample carrier in a contact-type or non-contact-type manner, and the acquisition module is connected to the analysis module by wire or wirelessly.

[0026] A gene sequencing method provided in the present application includes the following steps: loading a sequencing sample, loading the sequencing sample into a sample carrier; providing sequencing reagents, providing the sample with the reagents required for the sequencing reaction; collecting fluorescent signals, collecting fluorescent signals emitted by the sample; outputting sequence information, outputting base sequence information; the sample carrier includes a first substrate and a second substrate, the first substrate has a first attachment surface, the second substrate has a second attachment surface, the first attachment surface and the second attachment surface are arranged opposite to each other and are basically parallel, a swimming lane is formed between the first attachment surface and the second attachment surface, the swimming lane allows fluid to pass through or stay, and multiple samples are respectively attached to the first attachment surface and the second attachment surface.

[0027] The gene sequencing method provided in this application uses a sample carrier with a double-layer sample attachment surface, provides methodological guidance for the application of the double-layer sample carrier, and has the characteristics of high efficiency and high throughput.

[0028] Furthermore, the step of providing sequencing reagents and the step of collecting fluorescence signals are cycled several times.

[0029] Furthermore, the gene sequencing method also includes a step of preparing a sequencing sample, preparing a sample that can be loaded into the sample carrier for sequencing.

[0030] Furthermore, in the step of collecting the fluorescence signal, the fluorescence signal is collected on one side of the sample carrier.

[0031] Furthermore, in the step of collecting fluorescence signals, the fluorescence signals are collected on both sides of the sample carrier.

[0032] Furthermore, in the step of collecting the fluorescence signal, super-resolution imaging technology is used to collect the fluorescence signal.

[0033] Furthermore, the super-resolution imaging technology is structured illumination microscopy.

[0034] A sample carrier provided in the present application includes a first substrate and a second substrate, the first substrate having a first attachment surface, the second substrate having a second attachment surface, the first attachment surface and the second attachment surface are arranged opposite to each other and are basically parallel, a swimming lane is formed between the first attachment surface and the second attachment surface, the swimming lane allows fluid to pass through or stay, and the first attachment surface and the second attachment surface have attachment sites for placing samples.

[0035] The sample carrier provided in this application has two attachment surfaces, which fully utilizes the three-dimensional space. Compared with the single attachment surface of the prior art, it can carry more samples, greatly improves the utilization rate of the sample carrier, and can break through the limitations of mutual interference of fluorescent signals and improve sequencing throughput.

[0036] Furthermore, a vertical projection of the attachment position on the first attachment surface on the second attachment surface does not overlap with the attachment position on the second attachment surface, or a vertical projection of the attachment position on the second attachment surface on the first attachment surface does not overlap with the attachment position on the first attachment surface.

[0037] Furthermore, a vertical projection of the attachment position on the first attachment surface on the second attachment surface overlaps with the attachment position on the second attachment surface, or a vertical projection of the attachment position on the second attachment surface on the first attachment surface overlaps with the attachment position on the first attachment surface.

[0038] Furthermore, the attachment bits on the first attachment surface and / or the second attachment surface are arranged in a matrix.

[0039] Furthermore, the attachment locations on the first attachment surface and / or the second attachment surface are arranged in a staggered manner.

[0040] Furthermore, the attachment locations on the first attachment surface and / or the second attachment surface are randomly distributed.

[0041] Furthermore, the first substrate and / or the second substrate is made of a light-transmitting material.

[0042] Furthermore, the sample carrier has a plurality of swimming lanes.

[0043] The present application also provides another gene sequencing system, which includes a sample preparation module, a sample carrier, a fluid module, a collection module and an analysis module. The sample preparation module is used to prepare a sample that can be loaded into the sample carrier for sequencing. The sample carrier is used to carry the sequencing sample. The fluid module can provide the sample with the reagents required for the sequencing reaction. The collection module is used to collect the fluorescent signal emitted by the sample. The analysis module is used to analyze the fluorescent signal collected by the collection module and output sequence information.

[0044] This other gene sequencing system incorporates a sample preparation module and has the function of preparing samples, which simplifies the entire gene sequencing operation process and can improve sequencing efficiency and throughput.

[0045] Furthermore, the sample carrier is a single-layer sample carrier or a double-layer sample carrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 is a flow chart of the gene sequencing method provided in this application;

[0047] Figure 2 This is a block diagram of the gene sequencing system provided by this application;

[0048] Figure 3 This is a block diagram of an embodiment of the sample preparation module provided by this application;

[0049] Figure 4 This is a block diagram of an embodiment of the sample carrier provided by this application;

[0050] Figure 5 This is a block diagram of an implementation scheme of the acquisition module provided by this application;

[0051] Figure 6 is a schematic diagram of one embodiment of the sample carrier provided in this application;

[0052] Figure 7 yes Figure 6 A partial longitudinal cross-section of the sample carrier;

[0053] Figure 8 This is a schematic diagram of the first relative position relationship of samples on the two attachment surfaces of the sample carrier provided by the present application, wherein the collection module collects the fluorescence signal on one side of the sample carrier;

[0054] Figure 9 is a schematic diagram of the second relative position relationship of samples on the two attachment surfaces of the sample carrier provided by the present application, wherein the collection module collects the fluorescence signal on one side of the sample carrier;

[0055] Figure 10 This is a schematic diagram of the first relative position relationship of samples on the two attachment surfaces of the sample carrier provided by the present application, wherein the collection module collects the nearby fluorescence signals on both sides of the sample carrier;

[0056] Figure 11 This is a schematic diagram of the second relative position relationship of samples on the two attachment surfaces of the sample carrier provided by the present application, wherein the collection module collects the nearby fluorescence signals on both sides of the sample carrier;

[0057] Figure 12 This is a schematic diagram of the first relative position relationship of samples on the two attachment surfaces of the sample carrier provided by the present application, wherein the collection modules collect relatively distant fluorescence signals on both sides of the sample carrier;

[0058] Figure 13 This is a schematic diagram of the first arrangement of samples on a single attachment surface of the sample carrier provided in this application;

[0059] Figure 14 This is a schematic diagram of the second arrangement of samples on a single attachment surface of the sample carrier provided in this application;

[0060] Figure 15 This is a schematic diagram of the third arrangement of samples on a single attachment surface of the sample carrier provided in this application.

[0061] The following specific implementation methods will further illustrate this application in conjunction with the above-mentioned drawings. DETAILED DESCRIPTION

[0062] In order to make the purpose, technical solutions and advantages of this application more clear, the following further describes this application in detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to understand this application and are not intended to limit this application.

[0063] See also Figure 6 and Figure 7 The present application provides a sample carrier 9 for carrying a gene sequencing sample 29 (such as a DNA molecule), which includes a first substrate 27 and a second substrate 28. The first substrate 27 has a first attachment surface 30, and the second substrate 28 has a second attachment surface 31. The first attachment surface 30 and the second attachment surface 31 are arranged opposite to each other (facing each other, face to face) and are substantially parallel. A gap 32 (or "lane" in English) is formed between the first attachment surface 30 and the second attachment surface 31 to allow fluid to pass through (for example, along the Figure 6 The plurality of samples 29 are attached to the first attachment surface 30 and the second attachment surface 31 (e.g., Figure 7 As shown, Figure 7(This is a partial cross-sectional view of sample carrier 9, perpendicular to first attachment surface 30 and second attachment surface 31). This sample carrier 9 has two attachment surfaces, fully utilizing the three-dimensional space. Compared to the single attachment surface of the prior art, it can carry more samples, significantly improving sample carrier utilization, overcoming the limitations of mutual interference between fluorescent signals, and increasing sequencing throughput. For ease of reference, this document will also refer to this sample carrier 9 as a "double-layer sample carrier." The number of samples on each attachment surface may be in the hundreds of millions, and the size of a single sample may be only nanometers, which is not limited to the figure shown.

[0064] It should be noted that in the real world, it is difficult to achieve absolute parallelism between two planes. Due to the limitations of process conditions, the two planes can only be roughly parallel. Therefore, the term "basically parallel" is used above to describe the relative relationship between the first attachment surface 30 and the second attachment surface 31. This language is more rigorous and more in line with reality.

[0065] In some application scenarios, one or both of the first substrate 27 and the second substrate 28 can be made of a light-transmitting material, such as glass, plastic, rubber, or quartz. The light-transmitting material allows the fluorescence to pass through so that it can be collected.

[0066] The swimming lane 32 can be designed as a closed space or an open space according to the application scenario. If it is designed as a closed space, a closure can be set in places other than the area where the first attachment surface 30 and the second attachment surface 31 are attached to the sample, such as Figure 6 As shown, a sealant 26 is added between the first attachment surface 30 and the second attachment surface 31. If necessary, a hole can be opened in the enclosed space to the outside to serve as an inlet and outlet for the fluid. The hole can be opened on the first substrate and the second substrate (not shown) or on the sealant (not shown). If the design is an open space, then no sealant can be set between the first attachment surface 30 and the second attachment surface 31, or a sealant can be set only in some directions. In theory, the open space can be large enough to have no boundaries. As long as the distance between the first attachment surface 30 and the second attachment surface 31 is limited to a certain range, the fluid (liquid) can be confined to a specific area by relying on surface tension. The fluid (liquid) can also be confined to a specific area by designing the hydrophilicity and hydrophobicity of the attachment surface.

[0067] The orientations of the first attachment surface and the second attachment surface are not limited to the horizontal positions shown in the drawings, and they can be designed in any orientation according to the application scenario.

[0068] See also Figure 1In conjunction with the application of the above-mentioned sample carrier, the present application also provides a gene sequencing method 1, which includes step 2 of preparing a sequencing sample, step 3 of loading a sequencing sample, step 4 of providing a sequencing reagent, step 5 of collecting a fluorescent signal, and step 6 of outputting sequence information. Typically, the above steps are implemented in the following order: (1) preparing a sequencing sample; (2) loading a sequencing sample; (3) providing a sequencing reagent; (4) collecting a fluorescent signal; (5) outputting sequence information. If the sample 29 (such as a sequencing template) contains multiple bases to be tested, it is necessary to cycle the steps of "providing a sequencing reagent" and "collecting a fluorescent signal" several times until all bases are substantially detected.

[0069] Preparing sequencing samples refers to converting raw biological substances (such as blood, saliva, feces and tissues, etc.) or intermediate products into samples that can be attached to a sample carrier for sequencing after a series of treatments. This process may include multiple operational links, such as nucleic acid extraction, library amplification and quality control, etc. Each link can be completed by mechanical automation, by hand, or by a combination of the two methods. Intermediate products refer to the transitional products from the conversion of raw biological substances into the final sequencing samples. For example, in the prior art, interrupted DNA fragments will appear in the process of converting raw biological substances into the final sequencing samples. The interrupted DNA fragments are intermediate products. In this article, raw biological substances and intermediate products are collectively referred to as "raw materials."

[0070] Loading the sequencing sample refers to loading the prepared sequencing sample onto the sample carrier. The sample carrier referred to here is the sample carrier 9 provided in this application. During this process, the sample will adhere to the first attachment surface 30 and the second attachment surface 31.

[0071] Providing sequencing reagents refers to providing the reagents required for the sequencing reaction to the sample. For the present application, the reagents are introduced into the swimming lane 32 of the sample carrier 9 and undergo a biochemical reaction with the sample 29. Specifically, the reagents contain fluorescently labeled probes (such as dNTPs), and different probes complementarily bind to corresponding bases (also known as "synthesis reaction"), so that the sample carries the fluorescent label (fluorophore). It should be noted that in addition to the probes, the sequencing reagents may also contain other components required for sequencing, such as excision reagents, cleaning reagents, and purified water.

[0072] Collecting fluorescence signals refers to collecting the types of fluorescent markers bound to the sample. Optical devices, such as cameras, are usually used to collect fluorescence signals. Of course, the use of other devices that can collect fluorescence signals is not excluded. Some fluorescent markers require external light source excitation to emit fluorescent signals, while some fluorescent markers can generate fluorescent signals (which can be called "spontaneous emission") after binding to the sample, which depends on the selection of fluorescent marker materials.

[0073] Generally speaking, the process of continuously cycling through "providing sequencing reagents" and "collecting fluorescent signals" primarily involves synthesizing fluorescent probes, exciting fluorescent signals (if necessary), collecting fluorescent signals, and removing fluorescent groups. The reagent components, fluorescent labels (also referred to as "fluorescent markers," "fluorescent groups," etc.), and reaction processes described in this embodiment fall within the realm of biochemistry and are not described in detail in this application.

[0074] Outputting sequence information refers to outputting the base sequence information of the sample. Sequence information primarily represents the order of bases. This information is derived by integrating and analyzing the aforementioned fluorescence signals. This step involves complex information analysis techniques, which will not be explained in detail here.

[0075] The above-mentioned gene sequencing method 1 uses the sample carrier 9 provided in this application, and provides method guidance for the application of the sample carrier 9, which has the characteristics of high efficiency and high throughput.

[0076] As a variation of the above-mentioned gene sequencing method 1 (the first gene sequencing method), the present application also provides a second gene sequencing method, which includes step 3 of loading a sequencing sample, step 4 of providing a sequencing reagent, step 5 of collecting a fluorescent signal, and step 6 of outputting sequence information. Compared with the first gene sequencing method, the second gene sequencing method omits step 2 of preparing a sequencing sample, and the other steps and timing can be the same as the first gene sequencing method. The sequencing sample used in the second gene sequencing method can be ready-made and used directly, or prepared in a procedure other than this method.

[0077] See also Figure 2 The present application also provides a gene sequencing system 7, which includes a sample carrier 9, a fluid module 10, a collection module 11 and an analysis module 12.

[0078] The sample carrier 9 is used to carry the gene sequencing sample. In this embodiment, the sample carrier 9 provided above in this application is used. As another embodiment, the sample carrier may also be a sample carrier of other forms (such as a carrier with only one sample attachment surface, which for ease of description is also referred to as a "single-layer sample carrier" herein). The sample carrier can be either fixed or movable; it can be reusable or a consumable that needs to be replaced after a number of uses.

[0079] Fluidics module 10 can provide reagents required for sequencing reactions to samples, for example, providing reagents to sample 29 within sample carrier 9. This module can also implement the "providing sequencing reagents" step described above. If necessary, fluidics module 10 can also drain reagents, for example, removing waste liquid from the sample carrier. Alternatively, the fluidics module can also transport samples, loading sequencing samples into the sample carrier.

[0080] Acquisition module 11 can collect the fluorescence signal emitted by the sample. This module can implement the "collecting fluorescence signal" step described above. Acquisition module 11 can include a camera or other device capable of collecting fluorescence signals. The camera can be, for example, a CCD (Charge-Coupled Device) camera or a CMOS (Complementary Metal-Oxide-Semiconductor) camera, or, for example, an area array camera, a line array camera, or a TDI (Time Delay Integration) camera.

[0081] The analysis module 12 integrates and analyzes the fluorescence signals collected by the acquisition module 11 and outputs the sample's sequence information, i.e., the base sequence information of the sample. This module can also implement the "output sequence information" step described above. The acquisition module can transmit data to the analysis module, and the analysis module can receive data from the acquisition module. The sequence information output by the analysis module can be stored in an electronic memory, displayed on a display, transmitted to a computer server, or transmitted over a network.

[0082] In the gene sequencing system provided in the present application, the sample carrier 9 and the fluid module 10 can be connected in a contact manner (such as a pipeline connection) or in a non-contact manner (such as spraying or spraying the reagent onto the sample carrier through the air, and blowing away the waste liquid from the sample carrier with gas); the acquisition module 11 and the sample carrier 9 can be connected in a non-contact manner (such as there is a gap between the acquisition module 11 and the sample carrier 9) or in a contact manner (such as the acquisition module 11 is close to the sample carrier 9, and further, part or all of the acquisition module 11 is even integrated on the sample carrier 9 to form a whole with the substrate); the acquisition module 11 and the analysis module 12 can be connected in a wired signal manner or in a wireless signal manner.

[0083] The gene sequencing system 7 provided in this application provides an operating platform for the double-layer sample carrier 9, enabling it to perform gene sequencing. The system has the characteristics of high efficiency and high throughput. It should be noted that in addition to being applicable to double-layer sample carriers, the system is also applicable to single-layer sample carriers.

[0084] Furthermore, the gene sequencing system 7 provided in the present application may also include a sample preparation module 8. The sample preparation module 8 can convert the original biological material or intermediate product (raw material) into a sample that can be attached to a sample carrier for sequencing. This module can correspondingly implement the "preparing sequencing samples" step described above. As mentioned above, the process of preparing sequencing samples may include multiple operation links, and the sample preparation module 8 has the function of completing all operation links, or only has the function of completing some operation links.

[0085] The sample prepared by the sample preparation module 8 is transferred to the sample carrier, corresponding to the "loading sequencing sample" step described above. The sample can be transferred from the sample preparation module to the sample carrier by mechanical automation or by manual transfer. Of course, the two methods can also be mixed to achieve sample transfer. Mechanical automation methods, for example, can be transfer by a robotic arm or by a pipeline. As an embodiment, the fluid module also has the function of transferring samples. Manual methods, for example, can use a pipette to manually transfer samples. The gene sequencing system includes the sample preparation module 8, which makes the sequencing function more comprehensive and can complete more gene sequencing preparation processes.

[0086] The sample preparation module 8 is not limited to being an independent complete machine or component, but can also be a collection of multiple separate machines or components. Figure 3 As an embodiment, the sample preparation module 8 includes a plurality of sample preparation submodules (sample preparation submodules 1 to N, referenced as 13 to 16 ), each of which can implement different operation links.

[0087] A plurality of sample carriers 9 can be provided in the gene sequencing system 7, and each sample carrier 9 can also contain a plurality of sample sub-carriers, such as Figure 4 As shown. Each sample sub-carrier can contain an independent swimming lane. If a sample carrier contains multiple sample sub-carriers, it has multiple swimming lanes. Multiple sample sub-carriers can be physically separated or physically connected.

[0088] The gene sequencing system 7 provided in the present application is not limited to being an independent and complete machine. It can also be a collection of multiple separate machines or parts. For example, the sample preparation module 8 is a machine, and the sample carrier 9, fluid module 10, acquisition module 11 and analysis module 12 are integrated into another machine. These two machines are combined into the gene sequencing system 7, or the sample preparation module 8, sample carrier 9, fluid module 10, acquisition module 11 and analysis module 12 are integrated into one machine.

[0089] See also Figure 8, which is a schematic diagram of a first relative positional relationship of samples 29 on the two attachment surfaces of the sample carrier 9 provided in the present application, with the acquisition module 11 collecting fluorescence signals on one side of the sample carrier 9. In this first relative positional relationship, the vertical projection of the sample on one attachment surface onto the other attachment surface does not overlap with the sample on the other attachment surface. For example, the vertical projection of the sample 29 on the first attachment surface 30 onto the second attachment surface 31 does not overlap with the sample 29 on the second attachment surface 31. In other words, the vertical projection of the sample 29 on the second attachment surface 31 onto the first attachment surface 30 does not overlap with the sample 29 on the first attachment surface 30. The term "overlap" here can be understood as the intersection of the two figures, that is, the two figures have a common area.

[0090] The advantage of the first relative position relationship of the samples is that when the acquisition module 11 collects the fluorescence signal on one side of the sample carrier 9 (the side close to the first attachment surface 30 or the side close to the second attachment surface 31), the samples on the two attachment surfaces will not interfere with each other. Figure 8 In the figure, sample 29 on first attachment surface 30 and sample 29 on second attachment surface 31 are staggered, eliminating any obstruction. This reduces interference between the samples and the fluorescence. This positional relationship can, to a certain extent, lower the technical requirements for acquisition module 11 and minimize the difficulty of subsequent fluorescence signal analysis. In the figure, the fluorescence emitted by sample 29 on first attachment surface 30 is collected by the acquisition module through first substrate 27, while the fluorescence emitted by sample 29 on second attachment surface 31 is collected by the acquisition module 11 through first substrate 27. The dashed arrows in the figure indicate the general direction of fluorescence propagation. As long as the distance between the samples is within an appropriate range, strong interference between the fluorescence will not occur. Even if interference does occur, it will be within an acceptable range.

[0091] See also Figure 9 , which is a schematic diagram of a second relative positional relationship of samples on the two attachment surfaces of a sample carrier provided in the present application, with the acquisition module 11 acquiring fluorescence signals on one side of the sample carrier 9. In this second relative positional relationship, the vertical projection of the sample on one attachment surface on the other attachment surface overlaps with the sample on the other attachment surface. For example, the vertical projection of sample 29 on the first attachment surface 30 on the second attachment surface 31 overlaps with sample 29 on the second attachment surface 31. Alternatively, the vertical projection of sample 29 on the second attachment surface 31 on the first attachment surface 30 overlaps with sample 29 on the first attachment surface 30.

[0092] In the second relative position relationship of the samples, when the collection module 11 collects the fluorescence signal on one side of the sample carrier 9 , the samples on the two attachment surfaces will interfere with each other. Figure 9In the embodiment, the fluorescence emitted by the sample 29 on the first attachment surface 30 is collected by the collection module 11 through the first substrate 27, and the fluorescence emitted by the sample 29 on the second attachment surface 31 is blocked by the sample 29 on the first attachment surface 30. In this case, based on the diffraction phenomenon of light, the fluorescence emitted by the sample 29 on the second attachment surface 31 may still be collected by the collection module 11, but there are certain technical requirements for the collection module 11, and there are also certain requirements for the subsequent analysis of the fluorescence signal. However, these difficulties can be overcome, such as optimizing the hardware structure and analysis algorithm.

[0093] See also Figure 10 , which is a schematic diagram of the first relative position relationship of the samples on the two attachment surfaces of the sample carrier provided by this application, and the acquisition module 11 collects the nearby fluorescence signals on both sides of the sample carrier 9. Figure 8 The main difference shown is that the fluorescence emitted by the sample 29 on the first attachment surface 30 is collected by the collection module 11 through the first substrate 27, and the fluorescence emitted by the sample 29 on the second attachment surface 31 is collected by the collection module 11 through the second substrate 28. That is, the collection module 11 collects the fluorescence signal of the sample 29 on the attachment surface closest to it on the side of the first substrate 27 and the side of the second substrate 28 respectively. In this way, the light propagation is not blocked, which can reduce the difficulty of signal collection and differentiation and improve data quality.

[0094] See also Figure 11 , which is a schematic diagram of the second relative positional relationship of samples on the two attachment surfaces of the sample carrier 9 provided in this application. The collection module 11 collects nearby fluorescence signals on both sides of the sample carrier 9. The fluorescence emitted by the sample 29 on the first attachment surface 30 is collected by the collection module 11 through the first substrate 27, and the fluorescence emitted by the sample 29 on the second attachment surface 31 is collected by the collection module 11 through the second substrate 28. In other words, the collection module 11 collects the fluorescence signals of the sample on the attachment surface closer to the first substrate 27 and the second substrate 28, respectively. This ensures that light propagation is not obstructed, reducing the difficulty of signal collection and differentiation, and improving data quality.

[0095] See also Figure 12 , which is a schematic diagram of the first relative position relationship of the samples on the two attachment surfaces of the sample carrier 9 provided by this application, and the acquisition module 11 collects relatively far fluorescence signals on both sides of the sample carrier 9. Figure 10 The main difference is that the fluorescence emitted by the sample 29 on the first attachment surface 30 is collected by the collection module 11 through the second base 28, and the fluorescence emitted by the sample 29 on the second attachment surface 31 is collected by the collection module 11 through the first base 27, that is, the collection module 11 collects the fluorescence signals of the sample 29 on the attachment surface farther away from it on the side of the first base 27 and the side of the second base 28 respectively. Figure 8Similar to the embodiment shown, there is no obstruction during light propagation. As long as the distance between samples is within an appropriate range, there will be no strong interference between the fluorescence. Even if interference occurs, it will be within an acceptable range.

[0096] It should be noted that Figures 10 to 12 The collection modules 11 shown in the figure located on both sides of the sample carrier can be two independent modules or two sub-modules of a collection module 11. The collection module 11 is drawn as two independent modules in the figure mainly for the convenience of illustration, indicating that it collects fluorescence signals on both sides of the sample carrier 9, and does not limit the collection modules on both sides of the sample carrier to two independent collection modules 11.

[0097] In this application, the acquisition module 11 is not limited to being an independent complete machine or component, it can also be a collection of multiple separate machines or components, such as Figure 5 As shown, the acquisition module 11 may include multiple acquisition submodules (acquisition submodules 1 to N, referenced as 21 to 24 in the accompanying drawings). Each acquisition submodule may be a device with different functions. For example, different acquisition submodules acquire fluorescence signals of different colors (wavelengths), or may be components with different functional properties. Multiple acquisition submodules may acquire fluorescence signals from only one sample carrier 9, or may acquire fluorescence signals from multiple sample carriers 9. The acquisition module 11 may use a variety of technical principles to acquire fluorescence signals. For situations where the sample layout is densely packed, super-resolution imaging technology may be used, such as structured illumination microscopy (SIM) super-resolution imaging technology.

[0098] See also Figure 13 This application provides a first arrangement of samples 29 on a single attachment surface of a sample carrier 9. In this arrangement, multiple samples 29 are arranged in a matrix, also known as an "in-line arrangement." This arrangement provides a very regular arrangement of horizontal and vertical lines, making it easy to analyze. For example, after collecting fluorescence signals, it is easy to determine the physical location of each sample.

[0099] See also Figure 14 This application provides a second arrangement of samples 29 on a single attachment surface of a sample carrier 9. In this arrangement, multiple samples 29 are arranged in an array, with samples in adjacent rows staggered, also known as a "staggered arrangement." This arrangement can increase sample density and fully utilize the planar space of the attachment surface.

[0100] See also Figure 15The present application provides a third arrangement of samples 29 on a single attachment surface of a sample carrier 9, wherein multiple samples 29 are randomly distributed. This arrangement has the characteristics of low processing cost and relatively easy process.

[0101] It should be noted that the attachment surface usually needs to be provided with an attachment position for placing the sample. For example, the attachment position is chemically modified to form a specific chemical structure, so that when the sample passes through the attachment position, it can be captured and fixed at the position. For another example, the attachment position is charged, so when the sample passes through the attachment position, it can be captured and fixed at the position. Figures 13 to 15 The location of the sample shown is actually the location of the attachment site, and the three arrangements mentioned above are also the arrangements of the attachment site. (Similarly, Figures 8 to 12 The sample location shown is actually also the location of the attachment site. Of course, for the third arrangement, in another embodiment, the entire attachment surface serves as a large attachment site, and the sample is fixed at a random location as it passes through the attachment surface. An attachment site can be a concave surface, a convex surface, or an area flush with the attachment surface.

[0102] In objective reality, not every attachment position may be able to successfully place a sample, so some attachment positions will be vacant. Therefore, in this application, the distribution of samples according to the above three arrangement methods means that most samples are basically arranged in this way. The concept of this application cannot be circumvented on the grounds that individual attachment positions have not successfully placed samples or are vacant.

[0103] The first attachment surface 30 and the second attachment surface 31 can use the same sample arrangement mode or different sample arrangements. The sample arrangement mode can be selected entirely or partially on the same attachment surface, or a combination of the above arrangement modes can be used on the same attachment surface.

[0104] Please note that Figures 8 to 12 Shown is the relative position relationship between samples located on different attachment surfaces. Figures 13 to 15 The figure shows the relative position relationship between samples located on the same attachment surface. Please do not confuse them.

[0105] Figures 13 to 15 The arrangement of the samples on the attachment surface shown can be applied to both a double-layer sample carrier and a single-layer sample carrier.

[0106] The shape of the sample 29 shown in the figure is for illustration only and is not intended to limit its actual form. It can be in any form. For example, the sample 29 can be a single nucleic acid molecule, a cluster of multiple nucleic acid molecules, or a DNA nanoball (DNB).

[0107] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent replacements or improvements made within the spirit and principles of the present application should be included in the scope of protection of the present application.

Claims

1. A gene sequencing system comprising a sample carrier, a fluid module, a collection module, and an analysis module. The sample carrier is used to carry sequencing samples. The fluid module can provide the sample with reagents required for sequencing reactions. The collection module is used to collect fluorescent signals emitted by the sample. The analysis module is used to analyze the fluorescent signals collected by the collection module and output sequence information. The system is characterized in that: The sample carrier includes a first substrate and a second substrate, the first substrate has a first attachment surface, the second substrate has a second attachment surface, the first attachment surface and the second attachment surface are arranged opposite to each other and are basically parallel, a swimming lane is formed between the first attachment surface and the second attachment surface, the swimming lane allows fluid to pass through or stay, and multiple samples are respectively attached to the first attachment surface and the second attachment surface.

2. The gene sequencing system according to claim 1, characterized in that A vertical projection of the sample on the first attachment surface on the second attachment surface does not overlap with the sample on the second attachment surface, or a vertical projection of the sample on the second attachment surface on the first attachment surface does not overlap with the sample on the first attachment surface.

3. The gene sequencing system according to claim 1, characterized in that A vertical projection of the sample on the first attachment surface on the second attachment surface overlaps with the sample on the second attachment surface, or a vertical projection of the sample on the second attachment surface on the first attachment surface overlaps with the sample on the first attachment surface.

4. The gene sequencing system according to claim 1, wherein: The samples on the first attachment surface and / or the second attachment surface are arranged in a matrix.

5. The gene sequencing system according to claim 1, characterized in that: The samples on the first attachment surface and / or the second attachment surface are arranged in a staggered manner.

6. The gene sequencing system according to claim 1, characterized in that The samples on the first attachment surface and / or the second attachment surface are randomly distributed.

7. The gene sequencing system according to claim 1, characterized in that: The first attachment surface and the second attachment surface have attachment sites for placing samples, and the attachment sites on the first attachment surface and / or the second attachment surface are arranged in a matrix.

8. The gene sequencing system according to claim 1, characterized in that: The first attachment surface and the second attachment surface have attachment sites for placing samples, and the attachment sites on the first attachment surface and / or the second attachment surface are arranged in a staggered manner.

9. The gene sequencing system according to claim 1, characterized in that: The first attachment surface and the second attachment surface have attachment sites for placing samples, and the attachment sites on the first attachment surface and / or the second attachment surface are randomly distributed.

10. The gene sequencing system according to claim 1, characterized in that: The collection module collects the fluorescence signal on one side of the sample carrier.

11. The gene sequencing system according to claim 1, characterized in that: The collection module collects the fluorescence signals on both sides of the sample carrier.

12. The gene sequencing system according to claim 1, wherein: The acquisition module includes multiple acquisition submodules.

13. The gene sequencing system according to claim 1, characterized in that The acquisition module uses super-resolution imaging technology to acquire the fluorescence signal.

14. The gene sequencing system according to claim 1, characterized in that The acquisition module includes a CCD camera or a CMOS camera, or the acquisition module includes an area array camera, a line array camera or a TDI camera.

15. The gene sequencing system according to claim 1, characterized in that: The gene sequencing system includes a plurality of the sample carriers.

16. The gene sequencing system according to claim 1, characterized in that: The sample carrier includes a plurality of sample sub-carriers.

17. The gene sequencing system according to claim 1, characterized in that The gene sequencing system further includes a sample preparation module, which is used to prepare a sample that can be loaded into the sample carrier for sequencing.

18. The gene sequencing system according to claim 17, characterized in that: The sample preparation module includes a plurality of sample preparation submodules.

19. The gene sequencing system according to claim 17, wherein: The sample prepared by the sample preparation module is transferred to the sample carrier through a mechanical automation device or the fluid module.

20. The gene sequencing system according to claim 1, wherein: The sample carrier is associated with the fluid module in a contact-type or non-contact manner, the acquisition module is associated with the sample carrier in a contact-type or non-contact manner, and the acquisition module is connected to the analysis module by wire or wirelessly.

21. A gene sequencing method comprising the following steps: Loading sequencing samples into sample carriers; Providing sequencing reagents, providing the sample with reagents required for sequencing reaction; Collecting fluorescence signals, collecting fluorescence signals emitted by the sample; Output sequence information, output base sequence information; Its characteristics are: The sample carrier includes a first substrate and a second substrate, the first substrate has a first attachment surface, the second substrate has a second attachment surface, the first attachment surface and the second attachment surface are arranged opposite to each other and are basically parallel, a swimming lane is formed between the first attachment surface and the second attachment surface, the swimming lane allows fluid to pass through or stay, and multiple samples are respectively attached to the first attachment surface and the second attachment surface.

22. The gene sequencing method according to claim 21, characterized in that: The steps of providing sequencing reagents and collecting fluorescence signals are repeated several times.

23. The gene sequencing method according to claim 21, characterized in that The gene sequencing method further comprises a step of preparing a sequencing sample, wherein a sample is prepared that can be loaded into the sample carrier for sequencing.

24. The gene sequencing method according to claim 21, characterized in that In the step of collecting the fluorescence signal, the fluorescence signal is collected on one side of the sample carrier.

25. The gene sequencing method according to claim 21, characterized in that In the step of collecting fluorescence signals, the fluorescence signals are collected on both sides of the sample carrier.

26. The gene sequencing method according to claim 21, characterized in that In the step of collecting the fluorescence signal, super-resolution imaging technology is used to collect the fluorescence signal.

27. The gene sequencing method according to claim 26, characterized in that The super-resolution imaging technology is structured illumination microscopy.

28. A sample carrier, characterized in that: The invention comprises a first substrate and a second substrate, wherein the first substrate has a first attachment surface, and the second substrate has a second attachment surface. The first attachment surface and the second attachment surface are arranged opposite to each other and are substantially parallel to each other. A swimming lane is formed between the first attachment surface and the second attachment surface, and the swimming lane allows fluid to pass through or stay. The first attachment surface and the second attachment surface have attachment sites for placing samples.

29. The sample carrier according to claim 28, characterized in that A vertical projection of the attachment position on the first attachment surface on the second attachment surface does not overlap with an attachment position on the second attachment surface, or a vertical projection of the attachment position on the second attachment surface on the first attachment surface does not overlap with an attachment position on the first attachment surface.

30. The sample carrier according to claim 28, characterized in that A vertical projection of the attachment position on the first attachment surface on the second attachment surface overlaps with the attachment position on the second attachment surface, or a vertical projection of the attachment position on the second attachment surface on the first attachment surface overlaps with the attachment position on the first attachment surface.

31. The sample carrier according to claim 28, characterized in that The attachment sites on the first attachment surface and / or the second attachment surface are arranged in a matrix.

32. The sample carrier according to claim 28, characterized in that The attachment sites on the first attachment surface and / or the second attachment surface are arranged in a staggered manner.

33. The sample carrier according to claim 28, characterized in that The attachment sites on the first attachment surface and / or the second attachment surface are randomly distributed.

34. The sample carrier according to claim 28, characterized in that The first substrate and / or the second substrate are made of a light-transmitting material.

35. The sample carrier according to claim 28, characterized in that The sample carrier has a plurality of swimming lanes.

36. A gene sequencing system, characterized in that: It includes a sample preparation module, a sample carrier, a fluid module, a collection module and an analysis module. The sample preparation module is used to prepare samples that can be loaded into the sample carrier for sequencing. The sample carrier is used to carry sequencing samples. The fluid module can provide the samples with the reagents required for the sequencing reaction. The collection module is used to collect the fluorescent signal emitted by the sample. The analysis module is used to analyze the fluorescent signal collected by the collection module and output sequence information.

37. The gene sequencing system according to claim 36, characterized in that The sample carrier is a single-layer sample carrier or a double-layer sample carrier.

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