Sequencing system and liquid inlet method

Abstract

The invention provides a sequencing system and a liquid inlet method.The sequencing system comprises a sequencing chip, a reagent storage container, a liquid path system and a waste liquid storage system; a plurality of mutually isolated runners are arranged in the sequencing chip; the liquid path system comprises a plurality of multi-channel valves, a power pump and a parallel connector, and each multi-channel valve is provided with a public port, a blocking port, a plurality of connecting ports and a waste liquid port; the flow channels are divided into a plurality of groups, and the flow channels in the same group are connected in parallel through the parallel connectors and are communicated with one of the public ports; the number of the power pumps is the same as that of the flow channels in each group, each power pump is correspondingly provided with a section of rear-end pipeline, and the rear-end pipelines are communicated with valve heads of the power pumps; and one flow channel is arranged in each group of flow channels and is communicated with each section of the rear end pipeline. Through the technical scheme, the problem of reagent resource waste is reduced, and the image acquisition efficiency is improved.

Description

Technical Field

[0001] The present application relates to the technical field of gene sequencing, and in particular to a sequencing system and a liquid injection method. Background Art

[0002] In the gene sequencing system, the pump provides power and distributes the liquid reagents through the multi-channel rotary valve so that the target liquid reagents enter the flow channel of the predetermined sequencing chip; different flow channels can enter and exit the same or different fluids at the same or different times, and the liquid path system controls the fluid in and out of the flow channel to sequentially perform fluid transport, waiting and reaction, etc. Other systems of the instrument also perform their respective tasks in the above order.

[0003] The reaction of reagents in the flow channel only occurs in a very thin area, which is much thinner than the thickness of the flow channel. The effective components of the reagents involved in the reaction are usually less than 10%, or even less than 1%. After the reagents involved in the reaction in the flow channel of the flow cell flow out of the flow channel, they are discharged from the liquid path system to the waste liquid, which is a serious waste problem. Summary of the invention

[0004] The present application provides a sequencing system and a liquid injection method to reduce the waste of reagents in the gene detection process and save resources.

[0005] In a first aspect, the present application provides a sequencing system, including a sequencing chip, a reagent storage container, a liquid path system, and a waste liquid storage system;

[0006] The sequencing chip is provided with a plurality of mutually isolated flow channels;

[0007] The reagent storage container is provided with a plurality of containers for storing reaction reagents;

[0008] The liquid path system is used to connect the sequencing chip, the reagent storage container and the waste liquid storage system, and provide power for reagent transportation;

[0009] The fluid circuit system includes a multi-channel valve, a power pump and a parallel connector.

[0010] There are multiple multi-channel valves, each of which is provided with a common port, a blocking port, multiple connection ports and a waste liquid port;

[0011] The common port is in communication with the flow channel, the connection port is in communication with the reagent storage container, and the waste liquid port is in communication with the waste liquid storage system;

[0012] The common port is selectively connected to the blocking port, the plurality of connection ports and the waste liquid port;

[0013] The flow channels are divided into multiple groups, each group corresponds to one of the multi-channel valves, and each group of the flow channels includes multiple channels;

[0014] The flow channels in the same group are connected in parallel through the parallel connector and communicate with one of the common ports;

[0015] The number of the power pumps is the same as the number of the flow channels in each group, and each power pump is provided with a corresponding section of a rear end pipeline, and the rear end pipeline is communicated with the valve head of the power pump;

[0016] Each group of the flow channels is provided with a flow channel connected to each section of the rear end pipeline.

[0017] In a specific implementation scheme, there are four flow channels in total, with two channels forming a group;

[0018] The two flow channels of the same group are connected to the corresponding common port of the multi-channel valve through the parallel connector;

[0019] Each section of the rear-end pipeline is correspondingly provided with an adapter, and one of the flow channels in each group is connected in parallel through the adapter and communicated with the rear-end pipeline.

[0020] In a specific implementation manner, the adapter is a solenoid valve.

[0021] In a second aspect, the present application further provides a method for liquid injection into a sequencing system, based on any of the sequencing systems described above, the method comprising the following steps:

[0022] S1: adjusting the states of the multi-channel valve and the power pump to deliver the reagent to at least one of the flow channels in the first group of flow channels;

[0023] S2: adjusting the multi-channel valve, the power pump sucks liquid, and the reagent in the flow channel is transported from the flow channel to the corresponding rear-end pipeline, and another reagent is transported into the flow channel at the same time;

[0024] S3: adjusting the multi-channel valve and the power pump to deliver the reagent in the rear-end pipeline to the flow channel in the second group of flow channels connected to the same rear-end pipeline;

[0025] The first group of flow channels and the second group of flow channels are groups of flow channels corresponding to different multi-channel valves respectively.

[0026] In a specific implementation manner, in step S1, the reagent is delivered to all flow channels of the first group of flow channels;

[0027] After the reagents have reacted in all the flow channels of the first group of flow channels, an optical system is used to collect images of the interior of the first group of flow channels;

[0028] After the step S3 is completed and the reagent reacts in all the flow channels of the second group of flow channels, an optical detection system is used to collect images of the interior of the second group of flow channels.

[0029] In a specific embodiment, after the step S3 is completed, the second group of flow channels is supplemented with the same unreacted reagent as that in the second group of flow channels.

[0030] In a specific implementation scheme, the step S1 is to deliver the reagent into one of the flow channels of the first group of flow channels;

[0031] After the reagent is transported into one of the flow channels of the first group of flow channels and before the reagent reacts in the flow channel of the first group, the step S2 is performed, that is, in step S3, the unreacted reagent is transported from the rear end pipeline to a flow channel of the second group of flow channels connected to the same rear end pipeline;

[0032] After step S3 is completed, the reagents react simultaneously in the multiple flow channels corresponding to the same rear-end pipeline.

[0033] In a specific possible implementation scheme, a reagent is delivered into the flow channel for liquid injection, and after the liquid injection is completed and the reagent reacts in the flow channel, an optical detection system is used to collect images of the interior of the flow channel;

[0034] After completing the image acquisition, re-delivering the reagent into the corresponding flow channel;

[0035] Each of the flow channels continuously cycles through the process of liquid inlet-reaction-image acquisition;

[0036] The optical detection system collects images of the plurality of flow channels connected to the first power pump and collects images of the plurality of flow channels connected to the second power pump alternately.

[0037] In a specific possible implementation, in the steps S1 and S3, the corresponding solenoid valves are adjusted so that the rear-end pipelines are connected to different flow channels respectively.

[0038] In a specific implementation scheme, in step S1, the common port of the first multi-channel valve is adjusted to be connected to one of the connection ports connected to the reagent storage container, and the other multi-channel common ports are connected to the blocking port, and the power pump corresponding to each flow channel for transporting the reagent sucks liquid;

[0039] In step S2, the common port of the first multi-channel valve is adjusted to be connected to one of the connection ports connected to the reagent storage container, and the power pump that performs the liquid aspiration action in step S1 continues to aspirate liquid;

[0040] In the step S3, the common port of the first multi-channel valve is adjusted to be connected to the blocking port, and the common port of the second multi-channel valve is adjusted to be connected to the waste liquid port;

[0041] The first multi-channel valve is a multi-channel valve corresponding to the first group of flow channels;

[0042] The second multi-channel valve is a multi-channel valve corresponding to the second group of flow channels. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 An overall schematic diagram of the sequencing system provided in the embodiments of the present application;

[0044] Figure 2 A schematic diagram of the connection between the liquid circuit system provided in the embodiment of the present application and the sequencing chip, the reagent storage container, and the waste liquid storage container;

[0045] Figure 3 A schematic diagram of the connection between the flow channel, the multi-channel valve, and the power pump provided in the embodiment of the present application;

[0046] Figure 4 A schematic diagram of reagent delivery for one of the liquid injection methods provided in an embodiment of the present application;

[0047] Figure 5 A schematic diagram of delivering a reagent to a portion of a flow channel in another liquid injection method provided in an embodiment of the present application;

[0048] Figure 6 A schematic diagram of delivering reagent to another part of the flow channel in another liquid injection method provided in an embodiment of the present application. DETAILED DESCRIPTION

[0049] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below in conjunction with the accompanying drawings.

[0050] It should be noted that, unless otherwise defined, the technical terms or scientific terms used in one or more embodiments of this specification should be understood by people with ordinary skills in the field to which this disclosure belongs. The "first", "second" and similar words used in one or more embodiments of this specification do not indicate any order, quantity or importance, but are only used to distinguish different components. "Include" or "comprise" and similar words mean that the elements or objects appearing before the word cover the elements or objects listed after the word and their equivalents, without excluding other elements or objects. "Connect" or "connected" and similar words are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0051] To facilitate understanding of the sequencing system and liquid feeding method provided in the embodiment of the present application, its application scenario is first described. The sequencing system is used for gene sequencing work, including a sequencing chip, a chip platform, a reagent storage container, a liquid circuit system, etc., wherein the liquid circuit system is used for the transportation of sample libraries and reagents, transporting reagents to the sequencing chip for reaction, and transporting the reacted reagents to the waste liquid. Among them, amplification reaction and sequencing reaction occur in the sequencing chip, and image acquisition is performed by an image acquisition system to achieve gene sequencing. Exemplarily, a plurality of mutually isolated flow channels are provided in the sequencing chip, and the above reaction process is carried out in the flow channel, and the reaction only exists in a very thin area, the thickness of this area is much smaller than the flow channel thickness, that is, only part of the reagents transported to each flow channel participate in the reaction, and the remaining part is directly discharged to the waste liquid for collection after the reaction is completed; the effective ingredients of the reagents participating in the reaction are usually within 10%, or even within 1%, and the remaining reagents are all discharged to the waste liquid for centralized treatment, resulting in a more serious waste problem; and the cost of some of the reagents is high, which will also cause the problem of increased overall cost.

[0052] To this end, the present application provides a sequencing system and a liquid injection method to reduce reagent waste during sequencing, save resources, and reduce costs. The following is a detailed description of the system and the embodiments in conjunction with the specific drawings.

[0053] First reference Figure 1 , Figure 1The overall schematic diagram of the sequencing system is shown. The sequencing system includes a sequencing chip 10, a chip platform 20, a reagent storage container 30, a liquid circuit system 40, an optical detection system 50, a computer system 60 and a waste liquid storage container 70. Among them: the sequencing chip 10 is configured to provide a reaction area for amplification reaction and sequencing reaction; the chip platform 20 is configured to fix and support the sequencing chip 10; the reagent storage container 30 is configured to store one or more mixed sample libraries, one or more reagents; the liquid circuit system 40 is configured to controllably transport one or more mixed sample libraries, one or more reagents from the reagent storage container 30 to the sequencing chip 10, so as to perform amplification reaction and sequencing reaction in the sequencing chip 10, and controllably transport the waste liquid after the reaction from the sequencing chip 10 to the waste liquid storage container 70.

[0054] The optical detection system 50 is configured to excite and collect fluorescent signals after the sequencing reaction is completed, and generate fluorescent images based on the fluorescent signals; the computer system 60 is configured to obtain fluorescent images from the optical detection system 50 and identify the base sequence of the sample library based on the fluorescent images; the waste liquid storage container 70 is configured to store the waste liquid generated after the reaction.

[0055] refer to Figure 2 , Figure 2 The schematic diagram of the structure of the sequencing chip 10 is shown. The sequencing chip 10 is provided with one or more flow channels 11. When multiple flow channels 11 are provided, the flow channels 11 are isolated from each other. Each flow channel 11 has small holes at both ends for the inflow and outflow of fluids, wherein the fluids are biological samples and reaction reagents. For the sake of ease of description and understanding, the fluids are used as reagents as examples for illustration in this application.

[0056] refer to Figure 2 , Figure 2 The schematic diagram shows the matching relationship between the sequencing chip 10, the reagent storage container 30, the liquid circuit system 40 and the waste liquid storage container 70. The liquid circuit system 40 includes a pipeline connected between different components to provide a channel for reagent delivery, a switching structure for realizing the switching between different components of different pipelines, and a power pump 41 for providing power for the delivery of reagents. Specifically, the liquid circuit system 40 also includes a plurality of multi-channel valves 42, which are connected between the flow channel 11 and the reagent storage container 30 to realize the regulation of the delivery of different reagents to different flow channels 11.

[0057] Exemplarily, a plurality of flow channels 11 are provided, and the plurality of flow channels 11 are divided into a plurality of groups, each group corresponding to a multi-channel valve 42; wherein the multi-channel valve 42 is provided with a common port 421, a blocking port 422 and a plurality of connecting ports 423, and the common port 421 is selectively connected with the blocking port 422 and the plurality of connecting ports 423 through a rotary valve structure or an adjustable three-way structure.

[0058] A parallel connector 44 is provided between the multiple flow channels 11 of the same group and the corresponding multi-channel valve 42. The parallel connector 44 has multiple interfaces, one of which is connected to the common port 421 of the corresponding multi-channel valve 42, and the other multiple interfaces are respectively connected to the multiple flow channels 11 of the corresponding group, so as to realize the connection of the multiple flow channels 11 with the multi-channel valve 42 after being connected in parallel.

[0059] There are multiple reagent storage containers 30 for storing different reagents. The multiple connection ports 423 of the multi-channel valve 42 are respectively connected to different reagent storage containers 30. When transporting reagents, different connection ports 423 are adjusted to be connected to the common port 421 respectively, so that different reagents can be transported to the corresponding flow channel 11.

[0060] In addition, one of the connection ports 423 of each multi-channel valve 42 is connected to the waste liquid storage container 70, which is referred to as the waste liquid port 424 for ease of description and distinction; the reagents that have completed the reaction in the flow channel 11 are discharged into the waste liquid storage container 70 through the set waste liquid port 424 for storage, pending subsequent centralized processing.

[0061] refer to Figure 2 There are multiple power pumps 41, and the number of power pumps 41 is the same as the number of each group of flow channels 11; each power pump 41 is correspondingly provided with a rear end pipeline 43, one end of which is connected to a pump valve head port of the power pump 41, and the other end is connected to the flow channel 11, and the reagent is transported into the flow channel 11 by the power provided by the power pump 41. In addition, the other pump valve head port of the power pump 41 is connected to the waste liquid storage container 70, and the reagent after the reaction can be directly transported from the power pump 41 to the waste liquid storage container 70 for collection.

[0062] One end of the rear-end pipeline 43 connected to the flow channel 11 is connected to an adapter 45, and the adapter 45 is provided with multiple interfaces. The rear-end pipeline 43 is connected to multiple flow channels 11 through the adapter 45; specifically, one of the flow channels 11 in each group of flow channels 11 is connected to a section of the rear-end pipeline 43, and the above-mentioned multiple flow channels 11 are respectively connected to different ports of the adapter 45 to achieve parallel connection, and then connected to the corresponding rear-end pipeline 43. Among them, the number of ports provided on the adapter 45 is determined according to the number of groups of flow channels 11.

[0063] When the sequencing system is used for gene sequencing, the common port 421 of the multi-channel valve 42 corresponding to the flow channel 11 that needs to be supplied with liquid is adjusted to be connected with one of the connection ports 423 to deliver the corresponding reagent to the flow channel 11; the multi-channel valve 42 corresponding to the flow channel 11 that does not need to be supplied with liquid is adjusted to have the common port 421 connected with the blocking port 422, and the power pump 41 is operated to absorb liquid, and the reagent is delivered to the flow channel 11 through the multi-channel valve 42 for reaction.

[0064] After the reaction is completed, the power pump 41 performs a liquid suction action, so that the reagent in the flow channel 11 continues to be transported backward (i.e., in the direction of the power pump 41) and transported to the rear-end pipeline 43; in the process of transporting the reacted reagent in the flow channel 11 to the rear-end pipeline 43, the common port 42 of the multi-channel valve 42 corresponding to the flow channel 11 remains connected to one of the connection ports 423, and the reagent connected to the connection port 423 can be the same type of reagent as the above-mentioned reagent, or a new type of reagent, or a cleaning solution, buffer solution, etc.

[0065] The reacted reagents are transported to the rear-end pipeline 43, connecting the common port 421 of the multi-channel valve 42 corresponding to the above-mentioned channel 11 to the blocking port 422, connecting the common port 421 of another multi-channel valve 42 to the waste liquid port 424, and the power pump 41 is pushed back to transport the reagents to the channel connected to the same power pump 41 and corresponding to the next multi-channel valve 42.

[0066] In this way, the reagent can be reused. After the reagent completes a reaction in the flow channel 11, it is transferred through the rear-end pipeline 43 and transported to a different flow channel 11 connected to the same power pump 41 to react again, thereby improving the utilization rate of the reagent and reducing the problem of reagent waste.

[0067] refer to Figure 2 and Figure 3 There are four flow channels 11 in total, which are divided into two groups, each group having two flow channels; the parallel connector 44 is a three-way connector, two interfaces of which are connected to the two flow channels 11 in the same group, and the other structure is connected to the common port 421 of the corresponding multi-channel valve 42. The adapter 45 is also a three-way connector, one interface of which is connected to the rear-end pipeline 43, and the other two interfaces are respectively connected to the corresponding two flow channels 11.

[0068] In another embodiment, the adapter 45 is a solenoid valve, which can control the rear-end pipeline 43 to selectively connect with one of the flow channels 11. In this way, after the reagent reacts in one of the flow channels 11 and is transported to the corresponding rear-end pipeline 43, the adapter 45 is adjusted so that the rear-end pipeline 43 is connected to another flow channel 11 corresponding to the rear-end short circuit 43. At this time, the power pump 41 pushes back to transport the reagent in the rear-end pipeline 43 to another flow channel 11, avoiding the problem that the reagent in the rear-end pipeline 43 flows back to the original flow channel and is transported along the corresponding passage.

[0069] In addition, an embodiment of the present application further provides a liquid inlet method for a sequencing system, which is implemented based on the above-mentioned sequencing system, and the method includes the following steps:

[0070] S1: adjusting the states of the multi-channel valve 42 and the power pump 41 to deliver reagents to at least one of the flow channels 11 in the first group of flow channels 11;

[0071] S2: Adjust the multi-channel valve 42, the power pump 41 sucks the liquid, and the reagent in the flow channel 11 is transported from the flow channel 11 to the corresponding rear-end pipeline 43, and another reagent is transported into the flow channel 11 at the same time;

[0072] S3: adjusting the multi-channel valve 42 and the power pump 41 to deliver the reagent in the rear-end pipeline 43 to the flow channels 11 in the second group of flow channels 11 that are connected to the same rear-end pipeline 43;

[0073] The first group of flow channels 11 and the second group of flow channels 11 are groups of flow channels 11 corresponding to different multi-channel valves 42 , respectively.

[0074] In the first embodiment, step S1 is to transport the reagent into all the flow channels 11 of the first group of flow channels 11; after the reagent reacts in all the flow channels 11 of the first group of flow channels 11, the optical detection system 50 is used to capture images of the interior of the first group of flow channels 111;

[0075] After step S3 is completed, when the reagent has reacted in all the flow channels 11 of the second group of flow channels 11, the optical detection system 50 is used to capture images of the interior of the second group of flow channels 11; and after step S3 is completed, the second group of flow channels 11 can be supplemented with unreacted reagents of the same type as those therein according to actual needs.

[0076] Specifically, refer to Figure 4 , shows a schematic diagram of delivering reagents to the flow channel 11, wherein the process of reagent delivery is represented by a line segment with an arrow, and processes marked with the same numbers as I, II, III or IV are carried out simultaneously. For example, the two processes marked as I are for delivering reagents to two flow channels respectively, and the process is carried out simultaneously.

[0077] Specifically, a corresponding type of reagent is introduced into one group of flow channels 11, and a corresponding reaction process is carried out. After the reaction is completed, image acquisition is performed through the optical detection system 50; after the reaction in the group of flow channels 11 is completed and the image acquisition is completed, the reagent in the group of flow channels 11 is transferred to the rear-end pipeline 43, and then transported to each flow channel 11 of another group of flow channels 11 by the rear-end pipeline 43; thereafter, unreacted reagents are added to each flow channel 11 of the latter group of flow channels 11, and after the reaction is completed, image acquisition is performed.

[0078] By transferring through the rear-end pipeline 43, the reagent is transported in different flow channels 11, thereby achieving the reuse of the reagent and reducing the waste problem.

[0079] Specifically, the method includes the following steps: adjusting the common port 421 of the first multi-channel valve 42 among the plurality of multi-channel valves 42 to be connected to the reagent storage container 30, and the common ports 421 of the remaining multi-channel valves 42 are all connected to their respective blocking ports 422; multiple power pumps 41 simultaneously suck liquid to transport the reagent a from the corresponding reagent storage container 30 to each flow channel 11 of the first group of flow channels 11, and react in the flow channels 11. Among them, the first group of flow channels 11 is a group of flow channels 11 connected to the first multi-channel valve 42. For the sake of convenience, the second group of flow channels 11, the third group of flow channels 11, etc., which are defined later, are groups of flow channels 11 connected to different multi-channel valves 42, respectively.

[0080] After reagent a reacts in each flow channel 11 of the first group of flow channels 11, multiple power pumps 41 are controlled to continue to perform the liquid suction action, driving reagent a to be transported to the rear-end pipeline 43; in this process, the common port 421 of the first multi-channel valve 42 is kept connected with one of the connection ports 423, and the connection port 423 can be a port connected to reagent a, or a port connected to reagent b, or a port connected to a buffer solution or a cleaning solution. In the embodiment of the present application, the connection with reagent b is used as an example for explanation.

[0081] After the power pump 41 continues to aspirate liquid for a set time, the reacted reagent a is transported to the rear-end pipeline 43, and the process of transporting the reagent b to the flow channel 11 is completed; wherein, the set time is determined according to the aspiration power of the power pump 41 and the volume of the flow channel 11, and the corresponding relationship can be set before the sequencing system operates.

[0082] Then, the common port 421 of the first multi-channel valve 42 is adjusted to be connected to the blocking port 422, and the common port 421 of another multi-channel valve 42, for example, the second multi-channel valve 42, is connected to the waste liquid port 424; multiple power pumps 41 simultaneously perform a back-pushing action to drive the reagent a located in each rear-end pipeline 43 to be transported to the second group of flow channels 11, wherein the second group of flow channels 11 is a group of flow channels 11 connected to the second multi-channel valve 42.

[0083] In this way, the reagent a that has reacted in the first group of flow channels 11 can be transported to the second group of flow channels 11 for reaction again, rather than being directly discharged as waste liquid for collection and treatment, which can improve the utilization rate of resources and reduce waste; since the reagent reacts once in the flow channel 11, the proportion of the reagent that actually reacts is different, and the reagent a that has reacted in the second group of flow channels 11 can be reused later, that is, referring to the process of transferring the reagent a from the first group of flow channels 11 to the second group of flow channels 11, and then transferring the reagent a from the second group of flow channels 11 to the third group of flow channels 11, or even the fourth group of flow channels 11, the fifth group of flow channels 11, etc., to achieve more effective utilization of the reagent.

[0084] According to the different working scenarios of the sequencing system in actual situations, for example, the proportion of the reagent participating in the reaction each time is different, the number of times the reagent repeats the reaction is set to avoid the situation where the proportion of the reagent not participating in the reaction is small and affects the experimental results.

[0085] Of course, the reagent b delivered to the first group of flow channels 11, as well as the different types of reagents such as the reagent c and the reagent d that are subsequently input, can also be introduced by referring to the method of introducing and reusing the reagent a, which will not be described in detail here.

[0086] However, it is taken into consideration that as the number of reuses increases, the proportion of the reagent that has not participated in the reaction gradually decreases, which will have a certain impact on the reaction results; therefore, in the liquid inlet method of the embodiment of the present application, for each reagent that is reused, after it is transported to the flow channel 11 for secondary, tertiary, or other reactions, its purity and concentration are reduced, and new, identical reagents that have not reacted can be added thereto to ensure the accuracy of the experimental results.

[0087] Exemplarily, there are four flow channels 11, namely flow channel a, flow channel b, flow channel c, and flow channel d. The two multi-channel valves 42 are referred to as valve A and valve B, respectively. The two power pumps 41 are referred to as pump A and pump B, respectively. Among them, flow channel a and flow channel b are a group connected to valve A, which is the first group of flow channels 11; flow channel c and flow channel d are a group connected to valve B, which is the second group of flow channels 11. In addition, flow channel a and flow channel c are connected to pump A, and flow channel b and flow channel d are connected to pump B.

[0088] When working, the common port 421 of the regulating valve A is connected to the connecting port 423 connected to the reagent a, and the common port 421 of the valve B is connected to the blocking port 422; pump A and pump B simultaneously perform liquid suction action to transport the reagent a to the flow channel a and the flow channel b, and the reagent a reacts in the flow channel a and the flow channel b.

[0089] After the above reaction is fully completed, adjust the state of valve A so that its common port 421 is connected to the connecting port 423 connected to the reagent b, and valve B keeps the common port 421 connected to the sealing port 422. Pump A and pump B continue to perform the liquid suction action, driving the reagent a in the flow channel a and the flow channel b to be transported to the rear-end pipeline 423 corresponding to pump A and pump B. At the same time, the reagent b is transported to the flow channel a and the flow channel b, and the reagent b reacts in the flow channel 11.

[0090] Adjust the status of valve A and valve B so that the common port 421 of valve A is connected to the blocking port 422, and the common port 421 of valve B is connected to the waste liquid port 424; pump A and pump B push back at the same time to transport the reagent a temporarily stored in the rear-end pipeline 43 to the flow channel c and the flow channel d.

[0091] It should be explained that in the above process, the purpose of the power pump 41 pushing back is to transport the reagent a in the rear-end pipeline 43 to the flow channel c and the flow channel d. However, when the adapter 45 is a three-way, the power pump 41 pushes back, and the reagent a in the rear-end pipeline 43 may be transported to the flow channel a and the flow channel b. Specifically, taking the flow channel a as an example, the common port 421 of the valve A is connected with the sealing port 422, but the rear-end pipeline 43-flow channel a-flow channel b-flow channel d-valve B-waste liquid storage container 70 can still form a passage. However, since this transmission path will have greater resistance than the transmission from the rear-end pipeline 43 to the flow channel c, the probability of this transmission path appearing in actual work is low.

[0092] In one of the above embodiments, the adapter 45 is set as a solenoid valve, which can control the reagent transmission path by active adjustment; specifically, when the power pump 41 pushes back and transports the reagent A in the rear-end pipeline 43 to the flow channel c and the flow channel d, the two adapters 45 are adjusted so that the corresponding rear-end pipeline 43 is directly connected to the flow channel c and the flow channel d, and the passage from the rear-end pipeline 43 to the corresponding flow channel a and the flow channel b is closed. In this way, in the actual operation process, it is possible to avoid the situation where the reagent transmission direction deviates from the expected direction, which is conducive to more accurate control of the reagent delivery path.

[0093] At this time, the unreacted reagent b in flow channel a and flow channel b reacts therein, and the reagent a that has reacted once in flow channel c and flow channel d reacts therein, thereby realizing the reuse of reagent a and reducing waste. After reagent a has reacted in flow channel c and flow channel d, the common port 421 of valve B is connected to the waste liquid port 424, and pump A and pump B are pushed back to transport the reacted reagent a to the waste liquid storage container 70.

[0094] In addition, after the reagent a is pushed back from the rear end pipeline 43 to the flow channel c and the flow channel d, some unreacted reagent a can be added thereto, specifically as follows: the common port 421 of the adjustment valve B is connected to the connection port 423 connected to the reagent a, and the valve A still keeps the common port 421 connected to the blocking port 422, and the pump A and the pump B simultaneously suck liquid to deliver a certain amount of reagent a to the flow channel c and the flow channel d, and then react after mixing with the reacted reagent a. In this way, while realizing the reuse of the reagent, it can also reduce the impact on the experimental results and ensure the accuracy of the experiment.

[0095] After the reagent a in the flow channels c and d is discharged into the waste liquid storage container 70, the reagent b can be reused according to the above steps, and other reagents c, reagent d, etc. can also be reused according to the above steps.

[0096] In actual use, during the process of delivering different types of reagents to the flow channel 11 for reaction, the flow channel can be flushed with a cleaning liquid or a buffer to avoid mutual interference between different types of reagents; that is, reagent a is a reagent for reaction, and reagent b is a cleaning liquid or a buffer. In the embodiment of the present application, only the liquid feeding process is explained, and the specific types of reagents a and reagent b are not limited. Reagents a and reagents b can also be reagents of the same type. In actual practice, their types can be determined according to actual needs.

[0097] In the second embodiment, step S1 is to transport the reagent into one of the flow channels 11 of the first group of flow channels 111; after transporting the reagent into one of the flow channels 11 of the first group of flow channels 11 and before the reagent reacts in the flow channels 11 of the first group, step S2 is performed, that is, in step S3, the unreacted reagent is transported from the rear end pipeline 43 to a flow channel in the second group of flow channels 11 connected to the same rear end pipeline 43; after step S3 is completed, the reagent reacts simultaneously in multiple flow channels 11 corresponding to the same rear end pipeline 43.

[0098] During use, reagents are transported into the flow channel 11 for liquid injection. After the liquid injection is completed and the reagents react in the flow channel 11, images of the inside of the flow channel 11 are captured through the optical detection system 50. After the image capture is completed, reagents are re-transported into the corresponding flow channel 11. Each flow channel 11 continuously cycles through the process of liquid injection-reaction-image capture. The optical detection system 50 alternately performs the process of capturing images of the multiple flow channels 11 connected to the first power pump 41 and the process of capturing images of the multiple flow channels 11 connected to the second power pump 41.

[0099] Specifically, refer to Figure 5 and Figure 6 First, reagents are introduced into each flow channel 11 connected to the same power pump 41 in different groups of flow channels 11. For example, reagents are first transported to each flow channel 11 connected to the first power pump 41 in sequence. After the reagents are transported to each flow channel 11, the reagents react in the flow channels 11. After the reaction is completed, images are collected through the optical detection system 50.

[0100] In addition, reagents are transported into each flow channel 11 connected to the second power pump 411, and reacted therein. After the reaction is completed, images are collected by the optical detection system 50. The optical detection system 50 collects images of the multiple flow channels 11 connected to the first power pump 41 and the multiple flow channels 11 connected to the second power pump 41 alternately, that is, when collecting images of each flow channel 11 connected to the first power pump 411, each flow channel 11 connected to the second power pump 41 is in the process of liquid inflow or reagents reacting therein.

[0101] After the flow channel 11 connected to the first power pump 41 completes image acquisition, continue to input new reagents and repeat the process of liquid filling-reaction-image acquisition; at this time, the flow channels 111 connected to the second power pump 41 are in the state of liquid filling or reaction or the reagent has just completed the reaction. If the reagent has not reacted completely, image acquisition is directly performed after the reaction is completed.

[0102] As can be seen from the above, in the normal working process, each flow channel 11 can always maintain the cycle of liquid filling-reaction-image collection, and continue to fill the liquid after the image collection is completed; this avoids the problem of having to wait for a certain period of time before collecting images after a certain process is completed due to the different time arrangements for each process during the working process, thereby improving work efficiency.

[0103] Figure 5 , a schematic diagram of delivering a reagent to the flow channel 11 corresponding to the first power pump 41 is exemplarily shown, and the sequence of the reagent delivery process is represented by I, II, III, and IV; Figure 6 , a schematic diagram of conveying reagents to the flow channel 11 corresponding to the second power pump 41 is exemplarily shown, and the sequence of the reagent conveying process is indicated by I, II, III, and IV marked with circles on the periphery.

[0104] Compared with the liquid filling method of the prior art and the liquid filling method of the previous embodiment, in the liquid filling method of this embodiment, each flow channel 11 maintains the process of liquid filling-reaction-image collection continuously, thereby improving the image collection efficiency; it does not cause the problem of waiting for image collection for too long after the flow channel 11 has filled with liquid and reacted.

[0105] Specifically, the method comprises the following steps: S1: adjusting the states of the multi-channel valve 42 and the power pump 41 to deliver reagent a into one of the flow channels 11 of the first group of flow channels 11;

[0106] S2: adjusting the multi-channel valve 42 so that the flow channel 11 is connected to different reagent storage containers 30, delivering reagent b into the flow channel 11, and delivering reagent a to the rear-end pipeline 43 for temporary storage;

[0107] S3: adjusting the states of the multi-channel valve 42 and the power pump 41 to transport the reagent a temporarily stored in the rear-end pipeline 43 to another flow channel 11 connected to the same power pump 41 .

[0108] Then, repeat the above steps S2 and S3 to sequentially transport the reagents to the multiple flow channels 11 connected to the same power pump 41 until the reagents are transported to all the flow channels 11 connected to the power pump 41; wherein, the multiple flow channels 11 connected to the same power pump 41 can transport the same type of reagents or different types of reagents, and the specific type of reagent can be adjusted according to actual needs.

[0109] In each of the above steps, the specific operations for adjusting the power pump 41 and the multi-channel valve 42 are as follows, specifically in step S1: adjust the first multi-channel valve 42 among the multiple multi-channel valves 42 to a state where the common port 421 is connected to the reagent storage container 30, and the common ports 421 of the remaining multi-channel valves 42 are all connected to their respective blocking ports 422; adjust the first power pump 41 among the multiple power pumps 41 to absorb liquid, and deliver the corresponding reagent a to one of the channels in the first group of channels 11.

[0110] Step S2 specifically includes: adjusting the common port 421 of the first multi-channel valve 42 to connect to another different reagent storage container 30 , continuing the above-mentioned liquid aspiration process, delivering reagent b to the above-mentioned flow channel, and correspondingly, reagent a is transmitted to the rear-end pipeline 43 .

[0111] Specifically, step S3 includes: adjusting the common port 421 of the first multi-channel valve 42 to connect to the blocking port 422, and the common port 421 of another second multi-channel valve 42 to connect to the waste liquid port 424; the first power pump 41 is pushed back to transport the reagent a in the rear-end pipeline 43 to the flow channel 11 in the second group of flow channels 11 connected to the first power pump 41.

[0112] Afterwards, it can be determined whether new reagent a needs to be added to the flow channel 11 where reagent a is located. If necessary, the common port 421 of the second multi-channel valve 42 is adjusted to connect to the reagent storage container 30, and the first power pump 41 sucks liquid to complete the replenishment of reagent a.

[0113] After the reagent is transported, the reagent reacts in the flow channel 11. After the reaction is completed, image acquisition is performed through the optical detection system 50. During the process, the liquid filling and reaction process of each flow channel 11 corresponding to the second power pump 41 in each group of flow channels 11 can be carried out. For details, please refer to the liquid filling method of the flow channel 11 corresponding to the first power pump 41 in each group of flow channels 11 in this embodiment.

[0114] After the flow channel 11 corresponding to the second power pump 41 has completed the liquid inflow, the reaction is completed and the image is collected. When the image collection is completed, the flow channel 11 corresponding to the first power pump 41 is in a liquid inflow state, or in a state where the reagent is reacting in the flow channel 11, and after the reaction is completed, the image is collected through the optical detection system 50. In this way, the flow channels 11 corresponding to different power pumps 41 can form a cycle of liquid inflow, reaction and image collection.

[0115] Exemplarily, there are four flow channels 11, namely flow channel a, flow channel b, flow channel c, and flow channel d. The two multi-channel valves 42 are referred to as valve A and valve B, respectively. The two power pumps 41 are referred to as pump A and pump B, respectively. Among them, flow channel a and flow channel b are a group connected to valve A, which is the first group of flow channels 11; flow channel c and flow channel d are a group connected to valve B, which is the second group of flow channels 11. In addition, flow channel a and flow channel c are connected to pump A, and flow channel b and flow channel d are connected to pump B.

[0116] First, adjust the common port 421 of valve A to connect to the reagent storage container 30 corresponding to reagent a, and the common port 421 of valve B to connect to the blocking port 422, pump A sucks liquid to transport reagent a to flow channel a; switch the connection state of valve A so that its common port 421 is connected to the reagent storage container 30 corresponding to reagent b, pump A continues to suck liquid to transport reagent b to flow channel a, and at the same time, reagent a is transported to the rear-end pipeline 43.

[0117] Valve A is switched to the common port 421 to communicate with the blocking port 422, valve B is switched to the waste liquid port 424, pump A is pushed back, and reagent a in the rear end pipeline 43 is delivered to the flow channel c; at this time, the flow channel a contains reagent b, and the flow channel c contains reagent a, and then valve B is switched to the common port 421 to communicate with the reagent storage container 30 corresponding to reagent a, pump A sucks liquid, and replenishes part of reagent a into the flow channel c. Among them, whether new reagent a needs to be replenished into the flow channel c is determined according to the specific experimental situation, and whether to replenish should be based on ensuring the normal progress of the reaction.

[0118] The reagent reacts in flow channel a and flow channel c, and after the reaction is completed, image acquisition is performed through the optical detection system 50; in addition, during the process of acquiring images in flow channel a and flow channel c, flow channel b and flow channel d are in a liquid-filled state or a reagent-reacting state, and image acquisition is performed after the reagent reaction is completed, thus avoiding the problem of having to wait for a certain period of time before image acquisition can be performed.

[0119] The process of delivering reagents to flow channel b and flow channel d can refer to the above-mentioned process of delivering reagents to flow channel a and flow channel c; specifically, valve A is adjusted to the common port 421 connected to the reagent storage container 30, the common port 421 of valve B is connected to the blocking port 422, pump B sucks liquid, and the corresponding reagent is delivered to flow channel b; the common port 421 of valve A is connected to another reagent storage container 30, pump B continues to suck liquid, the latter reagent is delivered to flow channel b, and the former reagent is delivered to the rear-end pipeline 43; then valve A is switched to the common port 421 connected to the blocking port 422, the common port 421 of valve B is connected to the waste liquid port 424, pump B pushes back, pushes the reagent in the rear-end pipeline 43 back to flow channel d, and replenishes part of the former reagent to flow channel d. In other embodiments, reagents may no longer be replenished to flow channel d.

[0120] In this way, after the reagent reaction in flow channel a and flow channel c is completed, image acquisition is performed. During the image acquisition process, flow channel b and flow channel d are in a state of liquid intake or the reagent is reacting therein, and before the reagent reaction in flow channel b and flow channel d is completed, flow channel a and flow channel c complete the image acquisition process to avoid conflict with the subsequent image acquisition process of flow channel b and flow channel d, that is, the image acquisition process of flow channel a and flow channel c is performed alternately with the image acquisition process of flow channel b and flow channel d.

[0121] Compared with the liquid filling method in the prior art and another embodiment, the problem of mutual interference in the processes of liquid filling, reaction and image acquisition in different flow channels 11 is reduced, and the image acquisition efficiency is improved.

[0122] One or more embodiments of this specification are intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of one or more embodiments of this specification should be included in the scope of protection of this disclosure.

[0123] The above are only specific implementations of the present application, but the protection scope of the present application is not limited thereto. Any technician familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed in the present application, which should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

Claims

1. A sequencing system, characterized in that: Including sequencing chips, reagent storage containers, liquid circuit systems and waste liquid storage systems; The sequencing chip is provided with a plurality of mutually isolated flow channels; The reagent storage container is provided with a plurality of containers for storing reaction reagents; The liquid path system is used to connect the sequencing chip, the reagent storage container and the waste liquid storage system, and provide power for reagent transportation; The fluid circuit system includes a multi-channel valve, a power pump and a parallel connector. There are multiple multi-channel valves, each of which is provided with a common port, a blocking port, multiple connection ports and a waste liquid port; The common port is in communication with the flow channel, the connection port is in communication with the reagent storage container, and the waste liquid port is in communication with the waste liquid storage system; The common port is selectively connected to the blocking port, the plurality of connection ports and the waste liquid port; The flow channels are divided into multiple groups, each group corresponds to one of the multi-channel valves, and each group of the flow channels includes multiple channels; The flow channels in the same group are connected in parallel through the parallel connector and communicate with one of the common ports; The number of the power pumps is the same as the number of the flow channels in each group, and each power pump is provided with a corresponding section of a rear end pipeline, and the rear end pipeline is communicated with the valve head of the power pump; Each group of the flow channels is provided with a flow channel connected to each section of the rear end pipeline.

2. The sequencing system according to claim 1, characterized in that There are four flow channels in total, with two of them forming a group; The two flow channels of the same group are connected to the corresponding common port of the multi-channel valve through the parallel connector; Each section of the rear-end pipeline is correspondingly provided with an adapter, and one of the flow channels in each group is connected in parallel through the adapter and communicated with the rear-end pipeline.

3. The sequencing system according to claim 2, characterized in that The adapter is a solenoid valve.

4. A method for liquid inlet into a sequencing system, characterized in that: Based on the sequencing system according to any one of claims 1 to 3, the method comprises the following steps: S1: adjusting the states of the multi-channel valve and the power pump to deliver the reagent to at least one of the flow channels in the first group of flow channels; S2: adjusting the multi-channel valve, the power pump sucks liquid, and the reagent in the flow channel is transported from the flow channel to the corresponding rear-end pipeline, and another reagent is transported into the flow channel at the same time; S3: adjusting the multi-channel valve and the power pump to deliver the reagent in the rear-end pipeline to the flow channel in the second group of flow channels connected to the same rear-end pipeline; The first group of flow channels and the second group of flow channels are groups of flow channels corresponding to different multi-channel valves respectively.

5. The liquid inlet method of the sequencing system according to claim 4, characterized in that: In the step S1, the reagent is delivered to all the flow channels of the first group of flow channels; After the reagents have reacted in all the flow channels of the first group of flow channels, an optical system is used to collect images of the interior of the first group of flow channels; After the step S3 is completed and the reagent reacts in all the flow channels of the second group of flow channels, an optical detection system is used to collect images of the interior of the second group of flow channels.

6. The liquid inlet method of the sequencing system according to claim 5, characterized in that: After the step S3 is completed, the second group of flow channels is supplemented with the same unreacted reagent as that in the second group of flow channels.

7. The liquid inlet method of the sequencing system according to claim 4, characterized in that: The step S1 is to deliver a reagent into one of the flow channels of the first group of flow channels; After the reagent is transported into one of the flow channels of the first group of flow channels and before the reagent reacts in the flow channel of the first group, the step S2 is performed, that is, in step S3, the unreacted reagent is transported from the rear end pipeline to a flow channel of the second group of flow channels connected to the same rear end pipeline; After step S3 is completed, the reagents react simultaneously in the multiple flow channels corresponding to the same rear-end pipeline.

8. The liquid inlet method of the sequencing system according to claim 7, characterized in that: The reagent is transported into the flow channel for liquid injection. After the liquid injection is completed and the reagent reacts in the flow channel, an optical detection system is used to collect images of the interior of the flow channel; After completing the image acquisition, re-delivering the reagent into the corresponding flow channel; Each of the flow channels continuously cycles through the process of liquid inlet-reaction-image acquisition; The optical detection system collects images of the plurality of flow channels connected to the first power pump and collects images of the plurality of flow channels connected to the second power pump alternately.

9. The liquid inlet method of the sequencing system according to claim 4, characterized in that: In the steps S1 and S3, the corresponding solenoid valves are adjusted so that the rear-end pipelines are connected to different flow channels respectively.

10. The liquid inlet method of the sequencing system according to claim 4, characterized in that: In the step S1, the common port of the first multi-channel valve is adjusted to be connected to one of the connection ports connected to the reagent storage container, and the other multi-channel common ports are connected to the blocking port, and the power pump corresponding to each flow channel for conveying the reagent sucks liquid; In step S2, the common port of the first multi-channel valve is adjusted to be connected to one of the connection ports connected to the reagent storage container, and the power pump that performs the liquid aspiration action in step S1 continues to aspirate liquid; In the step S3, the common port of the first multi-channel valve is adjusted to be connected to the blocking port, and the common port of the second multi-channel valve is adjusted to be connected to the waste liquid port; The first multi-channel valve is a multi-channel valve corresponding to the first group of flow channels; The second multi-channel valve is a multi-channel valve corresponding to the second group of flow channels.

Images