Biological sample sequencing apparatus and operating method thereof

Abstract

According to one embodiment of the present disclosure, an operation method of a biological sample sequencing apparatus comprising a container array in which a plurality of containers are arranged according to a preset arrangement may comprise the steps of: a) controlling the temperature of a plurality of reagents stored in each of the plurality of containers to be constantly maintained at a set temperature for each reagent; b) introducing a reaction platform equipped with a biological sample library into one of the plurality of containers; c) removing the reaction platform from the one container after a predetermined time elapses; d) sequentially performing a container entry / exit operation of the reaction platform comprising the introduction operation and the removal operation on the plurality of containers, repeatedly up to the last container (nth container) of the container array; and e) repeating a sequencing cycle comprising steps b) to d) a predetermined number of times.

Description

Bio sample sequencing device and method of operation thereof

[0001] The present disclosure relates to a bio-sample sequencing device and a method of operating the same. Specifically, the present disclosure relates to a bio-sample sequencing device and a method of operating the same that significantly reduces the time required in the process of sequencing bio-samples, such as DNA and RNA.

[0002] DNA or RNA sequencing technology is performed by sequentially reacting multiple reagents with a library immobilized on the surface of a flow cell and analyzing the base sequence by detecting optical signals. The sequencing process is repeated approximately 300 times or more, and reagent transfer and temperature control are essential during this process.

[0003] Generally, reagent transfer is achieved by supplying the reagent into the flow cell through a needle and a tube using the negative and positive pressure of a syringe pump. Additionally, temperature control is achieved by regulating the temperature using a Peltier element located at the bottom of the flow cell. At this time, heat dissipation components such as a heat sink and a heat dissipation fan may be installed.

[0004] At this time, the reagent transfer and temperature control processes may take a significant amount of time, which leads to a problem where the overall sequencing speed is limited. In the reagent transfer method using a syringe pump, the transfer speed may be limited because it takes time for the reagent to completely fill the fluid channel of the flow cell from the container. Additionally, the method of variably controlling the temperature using a Peltier element requires repeated cooling and heating, so it may take time to reach the set temperature.

[0005] In summary, the sequencing process consists of four main steps: reagent transfer, temperature control, reagent reaction, and optical measurement. Among these, time delays occurring during the reagent transfer and temperature control steps can act as major bottlenecks that hinder the overall sequencing speed. Accordingly, in order to reduce the total time required for the sequencing process, it is necessary to shorten the reagent transfer and temperature control steps.

[0006] A method of operating a biosample sequencing device comprising a plurality of containers arranged according to a predetermined arrangement, disclosed as a technical means for achieving a technical task, may include: a) controlling the temperature of a plurality of reagents stored in each of the plurality of containers to maintain it at a constant temperature for each reagent; b) introducing a reaction platform equipped with a biosample library into one of the plurality of containers; c) removing the reaction platform from the one container after a predetermined time has elapsed; d) sequentially performing the container entry and exit operation of the reaction platform, consisting of the introduction operation and the removal operation, for the plurality of containers and repeating it up to the last container (n-th container) of the container array; and e) repeating the sequencing cycle consisting of steps b) to d) a predetermined number of times.

[0007] A bio-sample sequencing device disclosed as a technical means for achieving a technical task may include: a container array in which a plurality of containers for receiving reagents are arranged according to a preset arrangement; a temperature control unit for controlling the temperature of each reagent received within the plurality of containers; a reaction platform equipped with a bio-sample library where a reaction between the reagent and the bio-sample occurs; an actuator for controlling the movement of the reaction platform with respect to the plurality of containers; and a processor electrically connected to the temperature control unit and the actuator. The processor may control the temperature of the plurality of reagents stored in each of the plurality of containers to a set temperature for each reagent through the temperature control unit, insert the reaction platform equipped with the bio-sample library into one of the plurality of containers through the actuator, and remove the reaction platform from the one container through the actuator after a predetermined time has elapsed, and sequentially perform the container entry and exit operation of the reaction platform, consisting of the insertion operation and the removal operation, for the plurality of containers and repeat the process until the last container (n-th container) of the container array. The actuator can be controlled, and the actuator can be controlled to repeat the sequencing cycle, which consists of the container entry and exit operation performed n times, a predetermined number of times.

[0008] A computer-readable recording medium disclosed as a technical means for achieving a technical task has a program recorded thereon for performing the operation method of the aforementioned bio-sample sequencing device on a computer, and the method can be performed on a computer.

[0009] FIG. 1 is a schematic diagram illustrating a bio-sample sequencing device according to one embodiment of the present disclosure.

[0010] Figure 2 is a flowchart illustrating the overall operation of the bio-sample sequencing device shown in Figure 1.

[0011] Figure 3 is a flowchart illustrating the operation of acquiring a fluorescent signal and confirming a base sequence during the operation of the bio-sample sequencing device illustrated in Figure 1.

[0012] Figure 4 is a drawing illustrating an example of a container array applied to the bio-sample sequencing device shown in Figure 1.

[0013] Figure 5 is a drawing illustrating another example of a container array applied to the bio-sample sequencing device shown in Figure 1.

[0014] Figure 6 is a drawing illustrating another example of a container array applied to the bio-sample sequencing device shown in Figure 1.

[0015] FIGS. 7a to 7c are drawings illustrating examples of configurations for removing residual reaction reagents from a reaction platform during the sequencing process of a bio-sample sequencing device illustrated in FIG. 1.

[0016] In describing the present disclosure, technical details that are well known in the technical field to which the present disclosure belongs and are not directly related to the present disclosure are omitted. This is intended to convey the essence of the present disclosure more clearly without obscuring it by omitting unnecessary explanations. Furthermore, the terms described below are defined considering their functions within the present disclosure, and these definitions may vary depending on the intentions or practices of the user or operator. Therefore, their definitions should be based on the content throughout this specification.

[0017] For the same reason, some components in the attached drawings have been exaggerated, omitted, or schematically depicted. Additionally, the dimensions of each component do not entirely reflect their actual dimensions. Identical or corresponding components in each drawing have been assigned the same reference numbers.

[0018] The advantages and features of the present disclosure, and the methods for achieving them, will become clear by referring to the embodiments described below in detail together with the accompanying drawings. However, the present disclosure is not limited to the embodiments disclosed below but may be implemented in various different forms. The disclosed embodiments are provided to ensure that the disclosure of the present disclosure is complete and to fully inform those skilled in the art of the scope of the disclosure. An embodiment of the present disclosure may be defined according to the claims. Throughout the specification, the same reference numerals indicate the same components. Furthermore, in describing an embodiment of the present disclosure, if it is determined that a detailed description of a related function or configuration might unnecessarily obscure the essence of the present disclosure, such detailed description is omitted. Additionally, terms described below are defined considering their functions in the present disclosure, and these may vary depending on the intentions or conventions of the user or operator. Therefore, their definitions should be based on the content throughout the specification.

[0019] In one embodiment, each block of the flowcharts and combinations of the flowcharts may be executed by computer program instructions. Computer program instructions may be loaded onto a processor of a general-purpose computer, a computer for special purposes, or other programmable data processing equipment, and the instructions executed through the processor of the computer or other programmable data processing equipment may create means for performing the functions described in the flowchart block(s). Computer program instructions may also be stored in computer-available or computer-readable memory that can be directed toward the computer or other programmable data processing equipment to implement functions in a specific manner, and instructions stored in computer-available or computer-readable memory may produce a manufactured item containing instruction means for performing the functions described in the flowchart block(s). Computer program instructions may also be loaded onto a computer or other programmable data processing equipment.

[0020] Additionally, each block of the flowchart may represent a module, segment, or part of code containing one or more executable instructions for executing a specified logical function(s). In one embodiment, the functions mentioned in the blocks may occur out of order. For example, two blocks shown in succession may be executed substantially simultaneously or in reverse order depending on the function.

[0021] In one embodiment of the present disclosure, the term “part” used may refer to software or hardware components such as a Field Programmable Gate Array (FPGA) or an Application Specific Integrated Circuit (ASIC), and the “part” may perform a specific role. Meanwhile, the “part” is not limited to software or hardware. The “part” may be configured to reside in an addressable storage medium or may be configured to run one or more processors. In one embodiment, the “part” may include components such as software components, object-oriented software components, class components, and task components, as well as processes, functions, attributes, procedures, subroutines, segments of program code, drivers, firmware, microcode, circuits, data, databases, data structures, tables, arrays, and variables. Functions provided through a specific component or a specific “part” may be combined or separated into additional components to reduce their number. Additionally, in one embodiment, the “part” may include one or more processors.

[0022] Before describing specific embodiments of the present disclosure, the meanings of terms frequently used in this specification are defined.

[0023] A 'Bio Sample' may include a nucleic acid sequence composed of DNA (Deoxyribonucleic Acid) or RNA (Ribonucleic Acid). The bio sample covered in the present invention may undergo a preprocessing process before performing sequencing, and the process may include steps such as nucleic acid extraction, purification, fragmentation, reverse transcription, and adapter ligation.

[0024] A 'Bio Sample Library' refers to a collection of DNA or RNA fragments ready for sequencing. In other words, a Bio Sample Library loaded into a sequencing device is referred to as a 'Sequencing Library,' which can mean bio samples (DNA or RNA) that have undergone special processing (pretreatment) to enable sequencing. The library preparation process may include steps such as (i) fragmentation of the bio samples, (ii) attachment of adapters or barcodes, and (iii) PCR amplification or concentration adjustment. In the case of RNA sequencing, RNA samples are first converted into cDNA (complementary DNA) through a reverse transcription process, after which the library can be prepared in a manner similar to DNA sequencing. Finally, the completed library is loaded into the flow cell of the sequencing device to perform base sequence analysis.

[0025] "Reagent" refers to substances used in sequencing, encompassing a concept that includes various biochemical reagents utilized for the amplification of DNA or RNA libraries, base sequence reading, removal of reaction byproducts, and regulation of the reaction environment. Several reagents may be used during the sequencing process. First, base insertion and synthesis reagents may include fluorescently labeled nucleotides and DNA polymerase. As base insertion and synthesis reagents are applied to the library, a fluorescent signal may be generated when nucleotides are inserted. Fluorescence signal detection reagents stabilize the fluorescent signal and optimize signal intensity, allowing the base sequence to be read via a photodetector. After base reading is complete, fluorescent label removal reagents cleave and remove fluorescent labels and terminators using chemical or enzymatic methods to perform the next sequencing cycle. Washing and reaction environment reset reagents remove reaction byproducts after fluorescent label removal and optimize reaction conditions to ensure smooth subsequent base insertion. These processes are performed repeatedly, and each reagent plays a critical role in ensuring accurate base sequence reading.

[0026] A 'flow cell' refers to a structure in which DNA sequencing is performed while a biosample library is mounted and reagents are supplied sequentially. Flow cells can be designed based on microfluidic systems and may contain multiple reaction channels or microchannels that regulate fluid flow. Flow cells can be designed to precisely control the flow of reagents, induce biochemical reactions with biosamples, and ultimately detect and analyze fluorescent or electrical signals. Generally, flow cells are fabricated from glass, silicon, or polymer materials, and their surfaces may be chemically coated to immobilize or amplify DNA or RNA libraries. The role and structure of the flow cell may be designed differently depending on the sequencing method.

[0027] Embodiments of the present disclosure are described below with reference to the attached drawings so that those skilled in the art can easily implement them. However, the present disclosure may be embodied in various different forms and is not limited to the embodiments described herein.

[0028] Embodiments of the present disclosure will be described in detail below with reference to the drawings.

[0029] FIG. 1 is a schematic diagram illustrating a bio-sample sequencing device according to one embodiment of the present disclosure.

[0030] Referring to FIG. 1, a bio sample sequencing device (100) according to one embodiment may include a container array (110), a temperature control unit (120), a reaction platform (130), an actuator (140), a sensor unit (150), and a processor (not shown).

[0031] A container array (110) may include a plurality of containers (111, 112, 113, 114) for receiving reagents. Since different reagents are used depending on each step of the sequencing process, various reagents may be introduced into the plurality of containers (111, 112, 113, 114).

[0032] According to one embodiment, each reagent can be contained in a corresponding container so that various reagents do not mix with each other. That is, one reagent and one container have a one-to-one correspondence, and only one type of reagent can be contained in one container. Multiple reagents can be stored in each of the multiple containers (111, 112, 113, 114) that correspond to them.

[0033] In this case, the reagents stored in each container do not necessarily have to be different. That is, one type of reagent is stored in a single container, and the same type of reagent can be stored in each container.

[0034] A plurality of containers (111, 112, 113, 114) forming a container array (110) can be arranged according to a pre-set arrangement. As illustrated, the plurality of containers (111, 112, 113, 114) can be arranged in a line. Accordingly, a series of operations for sequentially performing the operation of introducing a reaction platform (130) into the container and removing the reaction platform (130) from the container for the plurality of containers (111, 112, 113, 114) can be facilitated, thereby shortening the time required for the entire sequencing process.

[0035] Meanwhile, multiple containers (111, 112, 113, 114) may be arranged in proportion to the number of reagents required in the sequencing process. As illustrated, a total of four containers are arranged in the container array (110), but the embodiment is not limited to the number of containers illustrated. Depending on the number of reagents required, the number of containers forming the container array (110) may increase or decrease.

[0036] Additionally, as illustrated, the volume of all containers forming the container array (110) is depicted as being the same, but the embodiment is not limited thereto. The volume of each container is determined by the size of the reaction platform (130) and the concentration of the reagent stored in the corresponding container, and may have different volumes for each container.

[0037] The temperature control unit (120) is configured to control the temperature of each reagent contained within a plurality of containers (111, 112, 113, 114). The temperature control unit (120) may include a plurality of heat sources. Each heat source can control the temperature of the reagent contained in each container.

[0038] At this time, since temperature control must be performed for each reagent, the temperature control unit (120) may include the same number of heat sources as the plurality of containers (111, 112, 113, 114) forming the container array (110). That is, each heat source can control the temperature of the reagent contained in the container corresponding to itself.

[0039] Each reagent contained in a plurality of containers (111, 112, 113, 114) can be controlled to a reagent-specific set temperature through a temperature control unit (120). In one example, the reagent-specific set temperature may be a temperature set by a user, and in another example, the reagent-specific set temperature may be a temperature stored in the bio-sample sequencing device (100) corresponding to the type of reagent. In this case, the processor can control the temperature control unit (120) to adjust the temperature of the reagent to a stored temperature according to the type of reagent.

[0040] Meanwhile, each reagent needs to be maintained at a constant temperature set for each reagent. By heating the reagents with the respective heat sources of the temperature control unit (120), the temperature of each reagent contained in the plurality of containers (111, 112, 113, 114) can be maintained at a constant temperature set for each reagent. According to this, when the temperature of the reagent reaches the temperature set for each reagent, there is no longer a need to raise or lower the temperature of the reagent.

[0041] At least from the moment the reagent temperature reaches the set temperature, the temperature of the heat source can be maintained at a constant level without change. In other words, the temperatures of both the heat source and the reagent can always be kept constant without the cumbersome process of heating or cooling the reagent during the sequencing process. Therefore, the 'time required for temperature control' can be effectively eliminated from the total time required for the sequencing process. This can significantly reduce the overall time required for the sequencing process.

[0042] In addition, generally, a Peltier element is used to perform heating and cooling of the reagent within a short time during the sequencing process; however, according to one embodiment, since there is no need to variably control the temperature of the reagent, such as by repeatedly heating and cooling the reagent, the heat source constituting the temperature control unit (120) is not necessarily limited to a Peltier element. The temperature control unit (120) may include various types of heat sources capable of maintaining a specific temperature for each reagent.

[0043] As illustrated, each heat source of the temperature control unit (120) may be positioned adjacent to each container. Specifically, each heat source may be positioned outside each container and may apply heat to the container. Since the heat source is positioned at the bottom of the container, heat generated from the heat source may be transferred to the reagent contained in the container through the bottom portion of the container. However, the position of the heat source in relation to the container is not limited to being positioned at the lower outer side of the container as illustrated. According to an embodiment, the heat source may be positioned inside the container to heat the reagent.

[0044] The reaction platform (130) is where the bio sample library is mounted. Sequencing can be performed after the bio sample library is mounted on the reaction platform (130). For example, the reaction platform (130) may include a flow cell.

[0045] According to one embodiment, the reaction platform (130) may include two reaction surfaces (131) facing each other and one or more contact surfaces (132) positioned to span across the two reaction surfaces. A biosample library may be mounted on each of the two reaction surfaces (131). That is, sequencing may substantially take place on the two reaction surfaces (131) of the reaction platform (130). One or more contact surfaces (132) are parts that come into contact with an actuator (140) to be described later. The actuator (140) may come into contact with one or more contact surfaces (132) and support the reaction platform (130).

[0046] As illustrated, the reaction platform (130) may have a rectangular shape. In this case, a surface having a relatively large area may be used as the reaction surface (131) to process a large amount of bio samples. There may be a total of four surfaces crossing two opposing reaction surfaces (131). However, as the actuator (140) contacts two of the four surfaces indicated by arrows, two contact surfaces (132) may be arranged facing each other. However, the shapes of the reaction platform (130) and the actuator (140) are not limited to those illustrated.

[0047] The actuator (140) may only come into contact with the contact surface (132) of the reaction platform (130) and not with the reaction surface (131). Since the actuator (140) does not interfere with the sequencing occurring on the reaction surface (131), the sequencing process can proceed more smoothly.

[0048] According to one embodiment, a reaction platform (130) can be introduced into a container forming a container array (110) by means of an actuator (140). The introduced reaction platform (130) can be removed from the container by means of an actuator (140) after a predetermined time has elapsed.

[0049] The insertion / removal operation of the reaction platform (130) can be performed sequentially for each container forming the container array (110). For example, a reaction platform (130) that is inserted into the first container (111) of the container array (110) and then removed from the first container (111) after a predetermined time may be inserted into a second container (112) adjacent to the first container (111). Afterward, the reaction platform (130) removed from the second container (112) may be inserted into and removed from the subsequent containers repeatedly. The insertion / removal operation of the reaction platform (130) may be repeated until it is performed for the last container (114), and in this way, one cycle of sequencing may proceed.

[0050] At this time, since the container already contains the reagent, when the reaction platform (130) is introduced into the container, the reagent inside the container can be processed by the reaction platform (130). Accordingly, there is no need to transfer the reagent to the reaction platform (130). In other words, the 'time required for transferring the reagent' can be effectively omitted from the total time required for the sequencing process. This can significantly reduce the total time required for the sequencing process.

[0051] In addition, in a general sequencing process, there is a high possibility of bubbles forming within the channel during the process of transferring fluid into the microfluidic channel of the reaction platform (130). At this time, the bubbles may cause interference when performing optical imaging for base sequence analysis after the reagent reaction.

[0052] However, the reaction platform (130) according to one embodiment does not require a separate channel, and sequencing is performed by placing the reaction platform (130) into a container and removing it from the container while the bio sample library is attached to the two reaction surfaces (131) of the reaction platform (130), thereby preventing the occurrence of bubbles. Accordingly, errors occurring during the process of acquiring and analyzing optical signals can be reduced.

[0053] The actuator (140) is a driving device for controlling the movement of the reaction platform (130) to move it to a desired position. The actuator (140) can perform physical movement using electric, hydraulic, pneumatic, or mechanical power. Depending on the driving method, the actuator (140) can be implemented as an electric, hydraulic, pneumatic, or mechanical type.

[0054] According to one embodiment, the actuator (140) has the primary purpose of moving the reaction platform (130) to a specific position. In this case, for precise movement control, the actuator (140) may be connected to a sensor, a processor, and a feedback system. Through this, the actuator (140) can accurately adjust the position of the reaction platform (130).

[0055] To control the movement of the reaction platform (130), the actuator (140) can support the reaction platform (130) through the contact surface (132). Contact between the actuator (140) and the reaction platform (130) can be maintained during the sequencing process.

[0056] The actuator (140) can align a reaction platform (130) on one of the multiple containers (111, 112, 113, 114) constituting the container array (110). As illustrated, the actuator (140) and the reaction platform (130) supported by it are located on the upper part of one container and are spaced apart from the container by a predetermined distance. That is, the reaction platform (130) can be in a position that is easy to insert into one container by the actuator (140).

[0057] The actuator (140) can introduce a reaction platform (130) into one container constituting the container array (110). Subsequently, the actuator (140) can remove the reaction platform (130) from the one container. Subsequently, the actuator (140) can move the reaction platform (130) to the next container adjacent to the one container. Specifically, the actuator (140) can align the reaction platform (130) on the next container. The above operations of the actuator (140) can be repeated until the reaction platform (130) reaches from the first container (111) to the last container (114).

[0058] When the actuator (140) removes the reaction platform (130) from the last container (114), one cycle of sequencing can be completed. The actuator (140) can move the reaction platform (130) to the first container (111) and align the reaction platform (130) on the first container (111). Then, by the actuator (140) introducing the reaction platform (130) into the first container (111), a second cycle can be performed.

[0059] Meanwhile, within one sequencing cycle, the process of confirming the base sequence of a bio sample mounted on a reaction platform (130) using a fluorescent substance may be performed. To this end, the bio sample sequencing device (100) may include a sensor unit (150).

[0060] The sensor unit (150) is configured to acquire a signal from the reaction platform (130). Specifically, the sensor unit (150) is configured to acquire data by detecting a fluorescent signal from a bio sample within the reaction platform (130) where sequencing has been performed. The sensor unit (150) can perform the function of detecting a fluorescent signal emitted from a fluorescent label attached to the bio sample, converting it into another signal, and transmitting it to a processor.

[0061] The sensor unit (150) may include an excitation light source for exciting a fluorescent label, an optical filter that selectively passes a fluorescent signal of a specific wavelength, a photodetector that detects the fluorescent signal and converts it into an electrical signal, and a signal processing module that processes the detected signal.

[0062] The sensor unit (150) can improve the detection accuracy of the detector by using a laser or LED to excite a fluorescent label and selectively separating the signal emitted from the fluorescent label through an optical filter.

[0063] An excitation light source can use a laser or LED to excite fluorescent labels attached to biological samples. After absorbing the excitation light, the fluorescent label can emit a fluorescent signal of a specific wavelength.

[0064] Optical filters separate excitation and emission light, allowing only light of specific wavelengths to pass through so that the emitted fluorescence signal can be accurately detected. Through this, optical filters can improve detection accuracy by eliminating unnecessary signals.

[0065] Fluorescence signals that have passed through an optical filter can be detected by a photodetector. The photodetector can convert the fluorescence signal into an electrical signal using a photomultiplier tube (PMT), a CCD sensor, or a CMOS sensor.

[0066] The Signal Processing Module can convert electrical signals into digital signals. Based on the converted digital signals, the Signal Processing Module can analyze and organize data regarding fluorescence intensity, fluorescence spectrum, and time-based signal variation. The organized data can be processed into a form that is easy for the processor to interpret through processes such as filtering and normalization. The processed data is transmitted to the processor, which can read the base sequence based on it.

[0067] Meanwhile, according to an embodiment, the function of the signal processing module may be performed by a processor. In this case, when an electrical signal generated by a photodetector is transmitted to the processor, the processor can determine the base sequence based on it.

[0068] As illustrated, the sensor unit (150) is shown acquiring a fluorescent signal from a bio sample in the reaction platform (130) after the reaction platform (130) is removed from the last container (114) of the container array (110). However, the time at which the fluorescent signal is acquired and the base sequence is confirmed is not limited to after the reaction platform (130) is removed from the last container (114). Depending on which of the multiple containers (111, 112, 113, 114) a reagent is stored, the time at which the fluorescent signal is acquired and the base sequence is confirmed may vary.

[0069] The processor is a central computing unit that is electrically connected to the temperature control unit (120), actuator (140), and sensor unit (150) to control the operation of each component and process data. The processor can be implemented as a Central Processing Unit (CPU), a Microcontroller Unit (MCU), a Digital Signal Processor (DSP), an Application-Specific Integrated Circuit (ASIC), etc.

[0070] The processor receives current temperature data from the temperature control unit (120) and can output a temperature control command to reach a set target temperature (e.g., a set temperature for each reagent). The processor can control the temperature control unit (120) to control the temperature of the reagent contained inside the container using a heat source. At this time, each heat source of the temperature control unit (120) can be controlled independently by the processor and maintained at a set target temperature.

[0071] The processor can control the movement of the reaction platform through the actuator (140). Specifically, the processor can control the actuator (140) so that the reaction platform (130) can move toward a plurality of containers forming the container array (110).

[0072] Additionally, when the sensor unit (150) detects a fluorescent signal from a bio sample library mounted on the reaction platform (130), the processor receives a signal and / or data corresponding to the fluorescent signal from the sensor unit (150) and analyzes it to read the base sequence information of the bio sample.

[0073] Consequently, the processor can coordinate the overall operation of the bio-sample sequencing device (100), such as controlling the temperature of the reagent, controlling the movement of the reaction platform (130), and analyzing the fluorescence signal, and can enable the sequencing process to proceed smoothly by analyzing the input data in real time and outputting an appropriate control signal.

[0074] Below, the operation of a bio-sample sequencing device into which a bio-sample library has been introduced will be described.

[0075] Figure 2 is a flowchart illustrating the overall operation of the bio-sample sequencing device shown in Figure 1.

[0076] In describing the operation of the bio sample sequencing device of FIG. 2 below, reference will be made to the components of the bio sample sequencing device (100) illustrated in FIG. 1.

[0077] Referring to FIG. 2, the operation of the bio sample sequencing device (100) can be started with a bio sample library introduced. The bio sample library can be mounted on each of the two opposing reaction surfaces (131) of the reaction platform (130).

[0078] In step S110, the bio sample sequencing device (100) can control the temperature of a plurality of reagents stored in each of a plurality of containers through a temperature control unit (120) to maintain the temperature of the reagents stored in each container at a constant temperature set for each reagent. Specifically, the processor can control the temperature control unit (120) to control the temperature of the reagents stored in each container to a temperature set for each reagent.

[0079] According to an embodiment, step S110 may be performed even before the bio sample library is mounted on the reaction platform (130). In this case, when the bio sample library is mounted on the reaction platform (130), the reagents in each container are already maintained at an optimal temperature for sequencing, so the bio sample sequencing device (100) can perform sequencing immediately.

[0080] Meanwhile, prior to the execution of step S120, the bio sample sequencing device (100) may support the reaction platform (130) via an actuator (140). Specifically, the actuator (140) may support the reaction platform by contacting one or more contact surfaces (132) that cross two reaction surfaces (131) of the reaction platform (130). This operation may be performed before step S110 is performed or after step S110 is performed.

[0081] In addition to step S110, when a bio sample library is mounted on the reaction surface (131) of the reaction platform (130) and an actuator (140) supports the reaction platform (130) through the contact surface (132) of the reaction platform (130), the processor determines that it is ready to perform sequencing on the bio sample, and step S120 can be performed.

[0082] In step S120, the bio sample sequencing device (100) can insert the reaction platform (130) equipped with the bio sample library into one of the containers (first container) forming the container array (110) via an actuator (140). For example, if sequencing has just started, the processor can control the actuator (140) to insert the reaction platform (130) into the first container (111) of the container array (110).

[0083] In step S130, the bio sample sequencing device (100) may remove the reaction platform (130) from the container via the actuator (140) after a predetermined time has elapsed. For example, the processor may control the actuator (140) to remove the reaction platform (130) from the first container (111). The predetermined time may mean a sufficient amount of time for the bio sample library mounted on the reaction platform (130) to interact with the reagent inside the container.

[0084] In step S140, the bio sample sequencing device (100) can determine whether there is another container in which the reaction platform (130) has not yet been introduced during the sequencing process. This determination can be performed by a processor.

[0085] In step S140, if it is determined that another container exists, steps S120 and S130 may be performed again for the next container placed adjacent to the container into which the reaction platform (130) was introduced. For example, under the control of a processor, the actuator (140) may perform the introduction / removal operation of the reaction platform (130) for the second container (112) adjacent to the first container (111).

[0086] At this time, the insertion operation of step S120 and the removal operation of step S130 can be combined and referred to as the 'container entry / exit operation of the reaction platform (130).' The processor can sequentially perform the container entry / exit operation, consisting of the insertion and removal operations of the reaction platform (130), for a plurality of containers (111, 112, 113, 114) through the actuator (140). The container entry / exit operation can be repeated up to the last container (n-th container) (114) of the container array (110). That is, steps S120 and S130 can be repeated until it is determined that there are no more containers.

[0087] At this time, since the multiple containers (111, 112, 113, 114) each contain different reagents, the reaction platform (130) can come into contact with multiple reagents until all entry and exit operations for the multiple containers (111, 112, 113, 114) are completed. Accordingly, multiple reagents can be processed sequentially in the bio sample library mounted on the reaction platform (130).

[0088] When the container entry / exit operation of the reaction platform (130) is performed with respect to the last container (114) of the container array (110), it may be determined in step S140 that no other containers exist. Accordingly, step S150 may proceed.

[0089] Meanwhile, according to an embodiment, the total number of containers into which the reaction platform (130) must be introduced before performing sequencing may be input into the processor. At this time, the total number of containers into which the reaction platform (130) must be introduced may be the same as the number of containers forming the container array (110).

[0090] According to this, step S140 may be omitted, and steps S120 and S130 may be repeated for each of the containers forming the container array (110) as many times as the total number of containers entered. When the repetition of steps S120 and S130 is finished, step S150 may proceed.

[0091] Steps S120 through S140 can form a single sequencing cycle. Generally, a sequencing cycle may consist of i) insertion of a fluorescently labeled nucleotide, ii) detection of a fluorescent signal, iii) removal of the fluorescent label and terminator, and iv) washing. That is, all four of the above processes can be performed while proceeding through steps S120 through S140.

[0092] In step S150, the bio sample sequencing device (100) can determine through a processor whether the sequencing cycle has reached a predetermined number of times. If it is determined that the sequencing cycle has reached the predetermined number of times, sequencing may be terminated. If it is determined that the sequencing cycle has not reached the predetermined number of times, steps S120 through S140 may be performed again. The bio sample sequencing device (100) may repeat the sequencing cycle consisting of steps S120 through S140 a predetermined number of times.

[0093] Meanwhile, among the four processes described above that constitute the sequencing cycle, the fluorescence signal detection process may have characteristics different from the other processes. While the other processes are performed by simply introducing a bio sample mounted on the reaction platform (130) into a container and bringing it into contact with a reagent, the fluorescence signal detection process can be performed by the connection between the sensor unit (150) and the processor, rather than by the introduction / removal operation of the reaction platform (130).

[0094] Below, we will describe the operation of obtaining a fluorescent signal from a bio sample mounted on a reaction platform (130) and confirming the base sequence.

[0095] Figure 3 is a flowchart illustrating the operation of acquiring a fluorescent signal and confirming a base sequence during the operation of the bio-sample sequencing device illustrated in Figure 1.

[0096] In describing the control operation of FIG. 3 below, reference will be made to the components of the bio-sample sequencing device (100) illustrated in FIG. 1.

[0097] Referring to FIG. 3, the fluorescent signal acquisition and base sequence verification operations can be performed between steps S130 and S140 of FIG. 2. That is, the fluorescent signal acquisition and base sequence verification operations can be performed after the container insertion operation of the reaction platform (130) for one container is performed and before the container insertion operation for the next container is performed.

[0098] Step S210 can be performed after Step S130 is performed. In Step S210, the bio sample sequencing device (100) can determine, through a processor, whether the preceding container into which the reaction platform (130) was introduced stores Fluorescent Signal Detection Reagents.

[0099] In this case, a fluorescence signal detection reagent may refer to a reagent that stabilizes the fluorescence signal and optimizes the signal intensity, enabling the base sequence to be read through a photodetector.

[0100] Specifically, fluorescent signal detection reagents may include fluorescence stabilizers and quenchers and optical detection reagents.

[0101] Fluorescence detection auxiliary reagents perform the role of signal stabilization and background signal reduction so that the fluorescence signal is clearly detected, and may include, for example, fluorescence stabilizers and quenchers. Photodetection reaction reagents are reagents that control reaction conditions to detect the fluorescence signal under optimal conditions, and may include, for example, tyramide signal amplification (TSA) reagents and optical buffer solutions.

[0102] In other words, fluorescent signal detection reagents are processed to enhance the quality of the fluorescent signal intended to be obtained from a bio-sample, and this can be distinguished from reagents processed to attach fluorescent substances or fluorescent labels to the bio-sample.

[0103] If it is determined that the preceding container into which the reaction platform (130) was introduced contains a fluorescent signal detection reagent, the processor determines that preparations for acquiring a fluorescent signal from the reaction platform (130) are complete, and step S220 may be performed. At this time, the processor may control the sensor unit (150) to acquire the fluorescent signal.

[0104] On the other hand, if the previous container into which the reaction platform (130) was introduced is not a container storing a fluorescent signal detection reagent, step S140 is performed so that the reaction platform (130) can be re-entered and exited from the container again.

[0105] Meanwhile, the reagent serving as the standard in step S210 is not limited to a fluorescent signal detection reagent. According to the embodiment, the processor may determine whether the preceding container stores a nucleic acid synthesis reagent or a washing reagent. Even in this case, if the processor determines that preparations for acquiring a fluorescent signal from the reaction platform (130) are complete, step S220 may be performed.

[0106] In step S220, the bio sample sequencing device (100) can acquire a signal from a bio sample or library mounted on a reaction platform (130) through a sensor unit (150). Specifically, the acquired signal may be a fluorescent signal emitted by a fluorescent substance (e.g., fluorescent label) attached to the bio sample after absorbing light, as previously mentioned.

[0107] In this case, step S220 may include the process of irradiating light onto a bio sample library mounted on a reaction platform (130) and the process of receiving light emitted from a fluorescent material attached to a bio sample inside the reaction platform (130).

[0108] For example, when a light source of the sensor unit (150) irradiates light onto a bio sample library mounted on a reaction platform (130), a light detector can receive light (e.g., a fluorescent signal) emitted from a fluorescent material attached to a bio sample inside the reaction platform (130). The fluorescence spectrum and fluorescence intensity of the fluorescent signal may vary depending on the attached base sequence during the sequencing process.

[0109] The photodetector can transmit a fluorescence signal to a processor. However, depending on the embodiment, the photodetector may generate another signal (e.g., an electrical signal) based on the fluorescence signal and transmit another signal corresponding to the fluorescence signal to the processor.

[0110] In step S230, the bio sample sequencing device (100) can determine the base sequence of the bio sample based on the signal acquired by the sensor unit (150) through a processor. For example, the signal acquired by the sensor unit (150) is a fluorescent signal, and the processor can receive the fluorescent signal acquired by the sensor unit (150) or receive an electrical signal or a digital signal generated by converting the fluorescent signal by the sensor unit (150). However, since the signal converted by the sensor unit (150) changes in correspondence with the fluorescent signal, the base sequence reading by the processor can be said to be fundamentally based on the fluorescent signal.

[0111] For example, when a processor receives a fluorescent signal, the processor can determine the base sequence attached to the bio sample during the sequencing process based on at least one of the fluorescence spectrum and fluorescence intensity of the fluorescent signal. Once the reading of the base sequence by the processor is finished, step S140 can be performed.

[0112] Below, we will describe the types and order of reagents sequentially processed on the reaction platform (130) by repeated container entry and exit operations within one sequencing cycle.

[0113] Figure 4 is a drawing illustrating an example of a container array applied to the bio-sample sequencing device shown in Figure 1.

[0114] Referring to FIG. 4, a bio sample sequencing device (100) according to one embodiment may include a container array (210) and a temperature control unit (220). Regarding the configuration and effects of the bio sample sequencing device (100), detailed descriptions that overlap with the above descriptions will be omitted, and the reaction platform (130) will be described with reference to the reference numeral shown in FIG. 1.

[0115] As described, the container array (210) may include four containers (211, 212, 213, 214). In this case, the temperature control unit (220) may include a heat source for each container. That is, the temperature control unit (220) includes four heat sources, and accordingly, the temperature for each container can be controlled.

[0116] The four containers (211, 212, 213, 214) can each contain different types of reagents. In this case, the multiple reagents stored in each container may include nucleic acid synthesis reagents, signal detection reagents, removal reagents, and washing reagents. In this case, the signal detection reagent may be the same as the fluorescent signal detection reagent described earlier.

[0117] Nucleic acid synthesis reagents may refer to the nucleotide incorporation and DNA synthesis reagents described above. Nucleic acid synthesis reagents may include fluorescently labeled nucleotides, each having a fluorescent label attached to it (A, T, G, C). Additionally, nucleic acid synthesis reagents may include DNA synthesis promoting reagents (DNA polymerase and buffer solutions). DNA synthesis promoting reagents may include DNA polymerase, an enzyme that inserts nucleotides into a DNA strand, and cofactors and buffers for reaction optimization.

[0118] Removal reagents may refer to fluorescent label and terminator removal reagents. Removal reagents can remove fluorescent substances (e.g., fluorescent labels) and terminators attached to biological samples.

[0119] Specifically, fluorescent labels can impart a specific fluorescent signal to each base (A, T, G, C) to enable the reading of the base sequence through optical detection, and terminators can ensure accurate base sequence reading by allowing only one nucleotide to bind at a time. Fluorescent labels and terminators are introduced to accurately read the base sequence, but they must be removed for the insertion of the next base. In particular, in sequencing using the Reversible Terminator Nucleotide (RT-NTP) method, the process of removing fluorescent labels and terminators may be essential for each cycle.

[0120] For example, cleavage reagents may include chemical cleavage reagents and enzymatic cleavage reagents. Chemical cleavage reagents are reagents that cleave fluorescent labels to restore nucleotides to their original state and may include reducing agents, acid solutions, etc. Enzymatic cleavage reagents are reagents for removing fluorescent labels and terminators using specific enzymes and may include hydrolases, nuclease-based cleavage enzymes, etc.

[0121] Washing reagents may refer to the aforementioned washing and buffering solutions. Washing reagents can be used during the washing process to remove remaining reaction byproducts after fluorescent labels and terminators have been removed.

[0122] For example, the washing reagents may include wash solutions and reconditioning buffers. The wash solutions can completely remove reaction by-products after fluorescent label removal to create an environment for the next cycle and may include buffer-based washing solutions, organic solvent-based washing solutions, etc. The reconditioning buffers can perform the role of resetting the reaction environment so that the next base insertion can proceed smoothly after washing the inside of the reaction platform (130) and may include pH buffer solutions, ion balance reagents, etc. Once the washing process is complete, the environment can be prepared so that the next nucleotide can be efficiently bound.

[0123] According to one embodiment, the four containers (211, 212, 213, 214) may include a first container (211) for receiving a nucleic acid synthesis reagent, a second container (212) for receiving a signal detection reagent, a third container (213) for receiving a removal reagent, and a fourth container (214) for receiving a first washing reagent.

[0124] The container entry and exit operation of the reaction platform (130) can be performed sequentially with respect to the first container (211), the second container (212), the third container (213), and the fourth container (214). Depending on the repeated container entry and exit operation of the reaction platform (130), the reaction platform (130) can sequentially come into contact with the reagents stored in each container. Accordingly, nucleic acid synthesis reagents, signal detection reagents, removal reagents, and washing reagents can be processed sequentially on the reaction platform (130).

[0125] During one sequencing cycle, the four types of reagents are processed sequentially, at least one base sequence of a bio sample mounted on the reaction platform (130) can be read, and preparation for performing the next sequencing cycle can be completed.

[0126] Figure 5 is a drawing illustrating another example of a container array applied to the bio-sample sequencing device shown in Figure 1.

[0127] Referring to FIG. 5, a bio sample sequencing device (100) according to one embodiment may include a container array (310) and a temperature control unit (320). Regarding the configuration and effects of the bio sample sequencing device (100), detailed descriptions that overlap with the above descriptions will be omitted, and the reaction platform (130) will be described with reference to the reference numeral shown in FIG. 1.

[0128] As illustrated, the container array (310) may include six containers (311, 312, 313, 314, 315, 316). In this case, the temperature control unit (320) may include a heat source for each container. That is, the temperature control unit (220) includes six heat sources, and accordingly, the temperature for each container can be controlled.

[0129] According to one embodiment, among the six containers (311, 312, 313, 314, 315, 316) of the container array (310), three containers (312, 314, 316) may store a washing reagent. At this time, the washing reagent may be divided into a first washing reagent that is processed on the reaction platform (130) after the removal reagent is processed on the reaction platform (130), and a second washing reagent that is processed on the reaction platform (130) after at least one of the nucleic acid synthesis reagent and the signal detection reagent is processed on the reaction platform (130). That is, the container array (310) shown in FIG. 5 may have a form in which two additional containers for accommodating the second washing reagent are added to the container array (210) shown in FIG. 4.

[0130] The first washing reagent and the second washing reagent are distinguished simply by the timing of processing on the reaction platform (130), and the two washing reagents may refer to substantially the same reagent. The timing of processing on the reaction platform (130) may vary depending on the arrangement of the container holding the reagent.

[0131] For example, when a base sequence is read via a fluorescent signal in one sequencing cycle, the removal of the fluorescent label and washing operations must be performed to prepare for the next sequencing cycle.

[0132] With this in mind, the removal reagent can be stored in the container immediately preceding the last container (316) (n-1th container) (315), and the first washing reagent can be stored in the last container (nth container) (316).

[0133] Accordingly, the removal reagent can be processed on the reaction platform (130) by the container entry / exit operation of the reaction platform (130) for the container immediately preceding the last container (315), and the first washing reagent can be processed on the reaction platform (130) by the container entry / exit operation of the reaction platform (130) for the last container (316). Accordingly, preparation for the next sequencing can be completed.

[0134] Meanwhile, the second washing reagent can be used to wash the reaction platform (130) to remove the reagent remaining on the reaction platform (130) according to the container entry and exit operation of the reaction platform (130).

[0135] If the container for the second washing reagent is referred to as the fifth container (312, 314), the fifth container (312, 314) can be placed next to at least one of the first container (first container) (311) for the nucleic acid synthesis reagent and the third container (second container) (313) for the signal detection reagent.

[0136] According to this, when at least one of the nucleic acid synthesis reagent and the signal detection reagent is processed in the reaction platform (130) according to the container entry / exit operation of the reaction platform (130), the second washing reagent can be processed in the reaction platform (130) by the container entry / exit operation of the reaction platform (130) that proceeds thereafter.

[0137] As illustrated, the fifth container (312, 314) may be placed between the first container (first container) (311) for receiving nucleic acid synthesis reagent, the third container (second container) (313) for receiving signal detection reagent, and the fifth container (third container) (315) for receiving removal reagent. That is, the second container (312) and the fourth container (314) may receive the second washing reagent.

[0138] According to this arrangement, the reaction platform (130) can be washed inside the second container (312) and the fourth container (314). Thus, the phenomenon in which the nucleic acid synthesis reagent of the first container (311) flows into the third container (313) through the reaction platform (130) and mixes with the signal detection reagent, or the nucleic acid synthesis reagent of the third container (313) flows into the fifth container (315) through the reaction platform (130) and mixes with the removal reagent, can be prevented.

[0139] According to this, the container array (310) can be utilized as is for the next sequencing task without any separate measures. As a result, the container array (310) can be used for a long time without a separate cleaning and reagent refilling process for the inside of the container.

[0140] Figure 6 is a drawing illustrating another example of a container array applied to the bio-sample sequencing device shown in Figure 1.

[0141] Referring to FIG. 6, a bio sample sequencing device (100) according to one embodiment may include a container array (410) and a temperature control unit (420). Regarding the configuration and effects of the bio sample sequencing device (100), detailed descriptions that overlap with the above descriptions will be omitted, and the reaction platform (130) will be described with reference to the reference numeral shown in FIG. 1. Meanwhile, for convenience of explanation, containers holding signal detection reagents and removal reagents have been omitted.

[0142] As illustrated, the container array (410) may include eight containers (411, 412, 413, 414, 415, 416, 417, 418). In this case, the temperature control unit (420) may include a heat source for each container. That is, the temperature control unit (420) includes eight heat sources, and accordingly, the temperature for each container can be controlled.

[0143] According to one embodiment, one or more nucleic acid synthesis reagents may be used depending on the sequencing method. As with the examples described above, the nucleic acid synthesis reagent may be contained in a container as a single reagent containing all bases (A, T, G, C), but as illustrated, the nucleic acid synthesis reagent may include four reagents according to the type of nucleic acid, and each of these may be contained in different containers among a plurality of containers.

[0144] Specifically, the first container (411) can accommodate only the A base synthesis reagent, the third container (413) can accommodate only the T base synthesis reagent, the fifth container (415) can accommodate only the G base synthesis reagent, and the seventh container (417) can accommodate only the C base synthesis reagent. According to this, the four reagents can be processed sequentially one by one on the reaction platform (130) according to the container entry and exit operation of the reaction platform (130).

[0145] However, in common to all examples of the container array (210, 310, 410), at least one of the one or more nucleic acid synthesis reagents may be stored in the first container (211, 311, 411) of the container array (210, 310, 410). Accordingly, at least one of the one or more nucleic acid synthesis reagents may be processed on the reaction platform (130) by the container entry / exit operation of the reaction platform (130) with respect to the first container (211, 311, 411) of the container array (210, 310, 410).

[0146] Simply put, among all the aforementioned reagents, the nucleic acid synthesis reagent can be processed first on the reaction platform (130). Since the sequencing of the bio sample begins with the reaction between the bio sample and the nucleic acid synthesis reagent, when sequencing begins, the reaction platform (130) is first introduced into the container containing the nucleic acid synthesis reagent, and accordingly, among the various types of reagents, the nucleic acid synthesis reagent can be processed first on the reaction platform (130).

[0147] Meanwhile, as illustrated, in the container array (410), a container for storing a washing reagent (412, 414, 416, 418) may be placed after the container (411, 413, 415, 417) for storing each nucleic acid synthesis reagent. Accordingly, the phenomenon of each nucleic acid synthesis reagent and other types of reagents mixing with each other within the container may be prevented. However, the placement of the washing reagent storage container and the relationship of placement with other containers are not limited to what is illustrated.

[0148] Below, examples of configurations for removing residual reagents from the reaction platform (130) without using a washing reagent will be described.

[0149] FIGS. 7a to 7c are drawings illustrating examples of configurations for removing reagents remaining on a reaction platform during the sequencing process of the bio-sample sequencing device illustrated in FIG. 1.

[0150] Referring to FIGS. 7a to 7c, a bio sample sequencing device (100) according to one embodiment may include a container array (510), a reaction platform (530), an actuator (540), and a reagent removal unit (560, 570, 580). Regarding the configuration and effects of the bio sample sequencing device (100), a detailed description that overlaps with the above description is omitted.

[0151] At this time, only one container forming the container array (510) is shown. However, the embodiment is not limited to the number of containers shown, and the description of the reagent removal unit (560, 570, 580) described later can be applied to all containers. Hereinafter, the 'container forming the container array (510)' shown in the drawing will be referred to as a container (510) by assigning a reference number.

[0152] The reagent removal unit (560, 570, 580) is configured to remove reagents remaining on the reaction platform (530) during the process of removing the reaction platform (530) from at least one of the plurality of containers (510).

[0153] Due to the arrangement of the reagent removal unit (560, 570, 580), the reaction platform (530) can be cleaned even if a container for receiving the cleaning reagent is not separately placed in the container array (510). Additionally, if both the reagent removal unit (560, 570, 580) and the container for receiving the cleaning reagent are arranged, the cleaning effect on the reaction platform (530) can be enhanced.

[0154] Referring to FIG. 7a, the reagent removal unit (560) may be a sprayer (560). In the process of removing the reaction platform (530) from the container (510), the sprayer (560) may spray a cleaning reagent, such as a cleaning solution, onto the reaction surface (531) of the reaction platform (530).

[0155] The sprayer (560) may be positioned at the inlet of the container (510). At this time, the container (510) may have a shape that narrows as it extends upward. Accordingly, the inlet of the container (510) located at the top of the container (510) may be relatively narrower than the bottom of the container (510). As a result, the sprayer (560) is positioned adjacent to the reaction platform (530) that is removed from the container (510), so that a sufficient amount of cleaning liquid can be supplied to the reaction platform (530). However, the shape of the container (510) is not limited to that depicted.

[0156] The reagent remaining on the reaction surface (531) can be washed away and removed by the cleaning solution sprayed from the sprayer (560). The removed reagent components can be returned to the interior of the container (510). At this time, the cleaning solution can also be returned to the interior of the container (510), but the cleaning solution used for cleaning may be composed of a substance that does not react with the reagent inside the container (510).

[0157] Referring to FIG. 7b, the reagent removal unit (570) may be a cleaning member (570). During the process of removing the reaction platform (530) from the container (510), the cleaning member (570) may come into contact with the reaction surface (531) of the reaction platform (530). The reagent remaining on the reaction surface (531) may be wiped away and removed by the cleaning member (570). At this time, some of the reagent may be absorbed by the cleaning member (570).

[0158] A cleaning member (570) may be placed at the entrance of the container (510). As the container (510) has a shape that narrows as it faces upward where the entrance of the container (510) is located, the cleaning member (570) may be positioned adjacent to the reaction platform (530) being removed from the container (510) to facilitate contact with the reaction surface (531).

[0159] The reagent remaining on the reaction surface (531) can be removed by being wiped or absorbed by the cleaning member (570). At this time, the cleaning member (570) may not have any significant effect on the bio sample mounted on the reaction surface (531). Therefore, sequencing of the bio sample mounted on the reaction platform (530) can proceed normally.

[0160] Meanwhile, according to an embodiment, in the process of removing the reaction platform (530) from the container (510), the cleaning member (570) may perform the function of a wiper that wipes the reaction surface (531) while moving along a predetermined trajectory.

[0161] Referring to FIG. 7c, the reagent removal unit (580) may be a vibrator (580). In the process of removing the reaction platform (530) from the container (510), the vibrator (580) may apply vibration to the reaction platform (530) to drop the reagent remaining on the reaction platform (530) into the interior of the container (510).

[0162] The vibrator (580) may be placed on a part of the actuator (540). During the process of removing the reaction platform (530) from the container (510), the vibrator (580) may be controlled to generate vibrations according to the commands of the processor. For example, when the vibrator (580) vibrates, vibrations may be transmitted through the actuator (540) to the reaction platform (530) that is in contact with the actuator (540). However, the location of the vibrator (580) is not necessarily limited to the actuator (540), and according to the embodiment, the vibrator (580) may be placed on a part of the reaction platform (530) that is not in contact with the reagent.

[0163] The reagent remaining on the reaction surface (531) can be removed from the reaction platform (530) by falling into the interior of the container (510) by vibrations generated by the vibrator (580). As the reagent is removed through the vibration method, the phenomenon of the reaction platform (530) or the reaction surface (531) coming into contact with other objects or substances and becoming contaminated can be prevented.

[0164] According to the embodiments described above, since there is no need to transfer reagents or control the temperature of reagents while sequencing is performed in the bio sample sequencing device (100), the time required for the sequencing process can be expected to be significantly reduced. In addition, the reaction platform of the bio sample sequencing device (100) does not require a separate microfluidic channel for transferring reagents, and since sequencing is performed by placing the reaction platform (130) into a container and removing it from the container while the bio sample library is attached to the two reaction surfaces, bubble generation is prevented, and thus the effect of reducing errors in the process of acquiring and analyzing optical signals can be expected.

[0165] A method of operating a bio-sample sequencing device according to one embodiment, comprising a plurality of containers arranged in a predetermined arrangement, may include: a) controlling the temperature of a plurality of reagents stored in each of the plurality of containers to maintain it at a constant temperature for each reagent; b) introducing a reaction platform equipped with a bio-sample library into one of the plurality of containers; c) removing the reaction platform from the one container after a predetermined time has elapsed; d) sequentially performing the container entry and exit operation of the reaction platform, consisting of the introduction operation and the removal operation, for the plurality of containers and repeating it up to the last container (n-th container) of the container array; and e) repeating the sequencing cycle consisting of steps b) to d) a predetermined number of times.

[0166] According to one embodiment, the plurality of reagents may include a signal detection reagent, and in steps b) and c), when one container accommodates the signal detection reagent, between steps c) and d), the method may further include the step of acquiring a signal from a bio sample mounted on the reaction platform, and the step of determining the base sequence of the bio sample based on the signal.

[0167] According to one embodiment, the signal may be a fluorescent signal emitted by a fluorescent substance attached to a bio sample after absorbing light, and the step of acquiring a signal from a bio sample mounted on a reaction platform may include the process of irradiating light onto a bio sample library mounted on the reaction platform and the process of receiving light emitted from a fluorescent substance attached to a bio sample inside the reaction platform.

[0168] According to one embodiment, the plurality of reagents may include a nucleic acid synthesis reagent, a signal detection reagent, a removal reagent, and a first washing reagent, and depending on the container entry and exit operation of the reaction platform that is repeatedly performed in step d), the nucleic acid synthesis reagent, the signal detection reagent, the removal reagent, and the first washing reagent may be sequentially processed on the reaction platform.

[0169] According to one embodiment, the removal reagent can be processed on the reaction platform by the container entry / exit operation of the reaction platform for the container immediately preceding the last container (n-1th container), and the first washing reagent can be processed on the reaction platform by the container entry / exit operation of the reaction platform for the last container (nth container).

[0170] According to one embodiment, the plurality of reagents may further include a second washing reagent, and when at least one of the nucleic acid synthesis reagent and the signal detection reagent is processed on the reaction platform according to the container entry / exit operation of the reaction platform, the second washing reagent may be processed on the reaction platform by the container entry / exit operation of the reaction platform that proceeds thereafter.

[0171] According to one embodiment, the plurality of reagents may include one or more nucleic acid synthesis reagents, and at least one of the one or more nucleic acid synthesis reagents may be processed on the reaction platform by the container entry / exit operation of the reaction platform with respect to the first container of the container array.

[0172] According to one embodiment, the nucleic acid synthesis reagent may include four reagents according to the type of nucleic acid, and the four reagents may be processed sequentially one by one on the reaction platform according to the container entry and exit operation of the reaction platform.

[0173] According to one embodiment, prior to step b), the method may further include the step of mounting the bio-sample library on each of two opposing reaction surfaces of the reaction platform, and the step of supporting the reaction platform by an actuator in contact with one or more contact surfaces that cross the two reaction surfaces of the reaction platform.

[0174] According to one embodiment, step c) may include a process of removing the reagent remaining on the reaction platform.

[0175] A bio-sample sequencing device according to one embodiment may include a container array in which a plurality of containers for receiving reagents are arranged according to a preset arrangement, a temperature control unit for controlling the temperature of each reagent contained within the plurality of containers, a reaction platform equipped with a bio-sample library where a reaction between the reagent and the bio-sample occurs, an actuator for controlling the movement of the reaction platform with respect to the plurality of containers, and a processor electrically connected to the temperature control unit and the actuator. The processor may control the temperature of the plurality of reagents stored in each of the plurality of containers to a set temperature for each reagent through the temperature control unit, insert the reaction platform equipped with the bio-sample library into one of the plurality of containers through the actuator, and remove the reaction platform from the one container through the actuator after a predetermined time has elapsed, and control the actuator to sequentially perform the container entry and exit operation of the reaction platform, consisting of the insertion operation and the removal operation, for the plurality of containers and repeat the process until the last container (n-th container) of the container array. The actuator can be controlled to repeat the sequencing cycle, which consists of the above container entry and exit operation performed n times, a predetermined number of times.

[0176] According to one embodiment, the system may further include a sensor unit for acquiring a signal from the reaction platform, and the plurality of reagents may include a signal detection reagent. When the processor determines that the one container into which the reaction platform is introduced contains the signal detection reagent, after removing the reaction platform from the one container, the processor may acquire a signal from a bio sample mounted on the reaction platform through the electrically connected sensor unit, and determine the base sequence of the bio sample based on the signal.

[0177] According to one embodiment, the signal may be a fluorescent signal emitted by a fluorescent substance attached to a bio sample after absorbing light, and the sensor unit may include a light source for irradiating light onto a bio sample library mounted on the reaction platform, and a photodetector for receiving light emitted from a fluorescent substance attached to a bio sample inside the reaction platform.

[0178] According to one embodiment, the plurality of reagents may include a nucleic acid synthesis reagent, a signal detection reagent, a removal reagent, and a first washing reagent, and the plurality of containers may include a first container for receiving the nucleic acid synthesis reagent, a second container for receiving the signal detection reagent, a third container for receiving the removal reagent, and a fourth container for receiving the first washing reagent, and the operation of receiving and receiving the containers of the reaction platform may proceed sequentially with respect to the first container, the second container, the third container, and the fourth container.

[0179] According to one embodiment, the removal reagent may be stored in the container immediately preceding the last container (the n-1th container), and the first washing reagent may be stored in the last container (the nth container).

[0180] According to one embodiment, the plurality of reagents may further include a second washing reagent, and the plurality of containers may further include a fifth container for receiving the second washing reagent, and the fifth container may be placed next to at least one of the first container and the second container.

[0181] According to one embodiment, the plurality of reagents may include one or more nucleic acid synthesis reagents, and at least one of the one or more nucleic acid synthesis reagents may be stored in the first container of the container array.

[0182] According to one embodiment, the reaction platform may include two reaction surfaces facing each other on which the bio sample library is mounted, and one or more contact surfaces crossing the two reaction surfaces, and the processor may control the actuator so that the actuator contacts the one or more contact surfaces to support the reaction platform.

[0183] According to one embodiment, the method may further include a reagent removal unit for removing the reagent remaining on the reaction platform during the process of removing the reaction platform from at least one of the plurality of containers.

[0184] A computer-readable recording medium according to one embodiment has a program recorded thereon for performing a method of operating a bio-sample sequencing device according to the above-described embodiment on a computer, so that the method can be performed on a computer.

[0185] Various embodiments of the present disclosure may be implemented or supported by one or more computer programs, and computer programs may be formed from computer-readable program code and stored on a computer-readable medium. In the present disclosure, “application” and “program” may represent one or more computer programs, software components, instruction sets, procedures, functions, objects, classes, instances, related data, or parts thereof suitable for implementation in computer-readable program code. “Computer-readable program code” may include various types of computer code, including source code, object code, and executable code. “Computer-readable medium” may include various types of media accessible by a computer, such as read-only memory (ROM), random access memory (RAM), hard disk drive (HDD), compact disc (CD), digital video disc (DVD), or various types of memory.

[0186] Additionally, a device-readable storage medium may be provided in the form of a non-transitory storage medium. Here, a 'non-transitory storage medium' is a tangible device and may exclude wired, wireless, optical, or other communication links that transmit transient electrical or other signals. Meanwhile, this 'non-transitory storage medium' does not distinguish between cases where data is stored semi-permanently and cases where it is stored temporarily. For example, a 'non-transitory storage medium' may include a buffer in which data is stored temporarily. A computer-readable medium may be any available medium accessible by a computer and may include both volatile and non-volatile media, as well as removable and non-removable media. A computer-readable medium includes media in which data can be stored permanently and media in which data can be stored and subsequently overwritten, such as rewritable optical discs or erasable memory devices.

[0187] According to one embodiment, the method according to the various embodiments disclosed herein may be provided by being included in a computer program product. The computer program product may be traded between a seller and a buyer as a product. The computer program product may be distributed in the form of a device-readable storage medium (e.g., compact disc read-only memory (CD-ROM)), or distributed online (e.g., download or upload) through an application store or directly between two user devices (e.g., smartphones). In the case of online distribution, at least a portion of the computer program product (e.g., a downloadable app) may be temporarily stored or temporarily created on a device-readable storage medium, such as the memory of a manufacturer's server, an application store's server, or a relay server.

[0188] The foregoing description of the present disclosure is for illustrative purposes only, and those skilled in the art will understand that modifications can be easily made to other specific forms without altering the technical spirit or essential features of the present disclosure. For example, suitable results may be achieved even if the described techniques are performed in a different order than described, and / or components such as systems, structures, devices, circuits, etc., described are combined or assembled in a form different from described, or replaced or substituted by other components or equivalents. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single unit may be implemented in a distributed manner, and components described as distributed may likewise be implemented in a combined form.

[0189] The scope of the present disclosure is defined by the claims set forth below rather than by the detailed description above, and all modifications or variations derived from the meaning and scope of the claims and equivalent concepts thereof should be interpreted as being included within the scope of the present disclosure.

Claims

1. A method of operating a biosample sequencing device comprising a container array in which a plurality of containers are arranged according to a preset arrangement, a) a step of controlling the temperature of a plurality of reagents stored in each of the plurality of containers to maintain it at a constant temperature set for each reagent; b) a step of introducing a reaction platform equipped with a bio-sample library into one of the plurality of containers; c) a step of removing the reaction platform from the one container after a predetermined time has elapsed; d) a step of sequentially performing the container entry / exit operation of the reaction platform, consisting of the above-described insertion operation and the above-described removal operation, for the plurality of containers and repeating the process until the last container (n-th container) of the container array; and e) a step of repeating the sequencing cycle consisting of steps b) to d) above a predetermined number of times; comprising a method.

2. In Paragraph 1, The above plurality of reagents includes a signal detection reagent, and In the above steps b) and c), When the above-mentioned container accommodates the signal detection reagent, Between step c) and step d) above, A step of acquiring a signal from a bio sample mounted on the above reaction platform; and A method further comprising the step of determining the base sequence of the bio sample based on the above signal.

3. In Paragraph 2, The above signal is a fluorescent signal emitted by a fluorescent substance attached to a bio-sample after absorbing light, and A method comprising the step of acquiring a signal from a bio sample mounted on the reaction platform, the step of irradiating light onto a bio sample library mounted on the reaction platform, and the step of receiving light emitted from a fluorescent substance attached to a bio sample inside the reaction platform.

4. In any one of paragraphs 1 through 3, The above plurality of reagents include a nucleic acid synthesis reagent, a signal detection reagent, a removal reagent, and a first washing reagent, and A method in which the nucleic acid synthesis reagent, the signal detection reagent, the removal reagent, and the first washing reagent are sequentially processed on the reaction platform according to the container entry and exit operation of the reaction platform that is repeatedly performed in step d) above.

5. In Paragraph 4, The above removal reagent is processed on the reaction platform by the container entry / exit operation of the reaction platform with respect to the container immediately preceding the last container (n-1th container), and A method in which the first washing reagent is processed on the reaction platform by the container entry / exit operation of the reaction platform with respect to the last container (n-th container).

6. In any one of paragraphs 1 through 3, The above plurality of reagents includes one or more nucleic acid synthesis reagents, and A method in which at least one of the above one or more nucleic acid synthesis reagents is processed in the reaction platform by the container entry / exit operation of the reaction platform with respect to the first container of the container array.

7. In any one of paragraphs 1 through 6, Prior to step b) above, The step of mounting the above bio-sample library on each of the two opposing reaction surfaces of the reaction platform; and A method further comprising the step of supporting the reaction platform by the actuator contacting one or more contact surfaces that cross the two reaction surfaces of the reaction platform.

8. An array of containers arranged according to a preset arrangement, wherein multiple containers for accommodating reagents are placed; A temperature control unit for controlling the temperature of each reagent contained within the plurality of containers above; A reaction platform equipped with a bio-sample library where a reaction between reagents and bio-samples takes place; An actuator for controlling the movement of the reaction platform with respect to the plurality of containers; and A processor electrically connected to the above temperature control unit and the above actuator; comprising The above processor is, The temperature of a plurality of reagents stored in each of the plurality of containers is controlled to a set temperature for each reagent through the above temperature control unit, and A reaction platform equipped with a bio-sample library is introduced into one of the plurality of containers through the above actuator, and After a predetermined period of time, the reaction platform is removed from the one container through the actuator, and The actuator is controlled to sequentially perform the container entry / exit operation of the reaction platform, which consists of the above-mentioned insertion operation and the above-mentioned removal operation, for the plurality of containers, and to repeat the operation until the last container (n-th container) of the container array. A bio-sample sequencing device that controls the actuator to repeat a sequencing cycle consisting of the above-mentioned container entry and exit operation, which is performed n times, a predetermined number of times.

9. In Paragraph 8, It further includes a sensor unit for acquiring a signal from the above-mentioned reaction platform, and The above plurality of reagents includes a signal detection reagent, and A bio sample sequencing device, wherein the processor determines that the one container into which the reaction platform is introduced contains the signal detection reagent, removes the reaction platform from the one container, acquires a signal from a bio sample mounted on the reaction platform through an electrically connected sensor unit, and determines the base sequence of the bio sample based on the signal.

10. In Paragraph 8, The above signal is a fluorescent signal emitted by a fluorescent substance attached to a bio-sample after absorbing light, and A biosample sequencing device comprising a sensor unit, a light source for irradiating light onto a biosample library mounted on the reaction platform, and a photodetector for receiving light emitted from a fluorescent substance attached to a biosample inside the reaction platform.

11. In any one of paragraphs 8 through 10, The above plurality of reagents include a nucleic acid synthesis reagent, a signal detection reagent, a removal reagent, and a first washing reagent, and The plurality of containers includes a first container for receiving the nucleic acid synthesis reagent, a second container for receiving the signal detection reagent, a third container for receiving the removal reagent, and a fourth container for receiving the first washing reagent. A bio-sample sequencing device in which the container entry and exit operations of the reaction platform are performed sequentially for the first container, the second container, the third container, and the fourth container.

12. In Paragraph 10, The above removal reagent is stored in the container immediately preceding the above last container (n-1th container), and A bio-sample sequencing device in which the first washing reagent is stored in the last container (n-th container).

13. In any one of paragraphs 8 through 10, The above plurality of reagents includes one or more nucleic acid synthesis reagents, and A biosample sequencing device in which at least one of the above one or more nucleic acid synthesis reagents is stored in the first container of the container array.

14. In any one of paragraphs 8 through 13, The above reaction platform comprises two reaction surfaces facing each other on which the bio sample library is mounted, and one or more contact surfaces crossing the two reaction surfaces. A biosample sequencing device, wherein the processor controls the actuator so that the actuator contacts one or more contact surfaces to support the reaction platform.

15. A computer-readable recording medium having a program recorded thereon for performing the method of any one of paragraphs 1 through 7 on a computer.

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