Automated sample preparation system

Abstract

An automatic sample preparation system comprises a consumable storage module, a liquid drop generation module, a biochemical reaction module, a quantification module and a scheduling transfer module, and the consumable storage module is used for storing consumables; the droplet generation module is used for generating micro-droplets containing biological samples; the biochemical reaction module comprises a plurality of biochemical reaction devices, and each biochemical reaction device is used for executing one or more sub-processes in gene sequencing sample preparation; the quantification module is used for quantitatively detecting the concentration of a product sample obtained in the sample preparation process; the dispatching and transferring module is used for grabbing and transferring the consumables and the samples and moving among all the positions along a preset path to complete transferring of the consumables and / or liquid. According to the automatic sample preparation system provided by the invention, a one-button single cell automatic sample preparation process is realized, a library sample for gene sequencing can be output after the sample is input, human intervention is reduced, manual operation errors are reduced, manpower is saved, the cost is reduced, and the result is more accurate and reliable.

Description

Technical Field

[0001] The present application relates to the technical field of biochemical substance analysis, and in particular to an automated sample preparation system. Background Art

[0002] Single-cell library construction is a key step in single-cell omics research, enabling the study of biological processes such as gene expression and epigenetic changes at the level of individual cells. The development of this technology has promoted our understanding of life processes, especially in fields such as developmental biology, tumor biology, and neuroscience.

[0003] Currently, single-cell library construction usually requires multiple separate instruments to complete the entire library construction process. There is no highly automated solution, especially the quantification of intermediate product concentrations, which is cumbersome and requires pausing the process and manually removing the intermediate product for quantitative testing. In addition, the throughput is low and the equipment is expensive. Utility Model Content

[0004] In view of this, in order to solve at least one of the above technical problems, it is necessary to propose an automated sample preparation system.

[0005] An embodiment of the present application provides an automated sample preparation system, comprising: a main housing and a consumables storage module, a droplet generation module, a biochemical reaction module, a quantification module, and a scheduling and transfer module integrated in the main housing, wherein the consumables storage module is configured to store consumables; the droplet generation module is configured to generate microdroplets containing biological samples; the biochemical reaction module includes a plurality of biochemical reaction devices, each of which is used to execute one or more sub-processes in a gene sequencing sample preparation process; the quantification module is configured to quantitatively detect the concentration of a product sample obtained in the gene sequencing sample preparation process; the scheduling and transfer module is configured to grab the consumables stored in the consumables storage module, and the scheduling and transfer module is further configured to move along a predetermined path between the consumables storage module, the droplet generation module, the biochemical reaction module, and the quantification module to complete the transfer of the consumables and / or liquids.

[0006] In some possible embodiments, the consumable storage module includes multiple consumable storage positions to store multiple consumables, and the multiple consumable storage positions include: a sample storage position for storing sample containers loaded with biological samples; a reagent storage position for storing reagent containers loaded with reagents required for biochemical reactions; a tip storage position for storing tips for transferring the biological samples or the reagents; a carrier plate storage position for storing chip carriers required for droplet generation; a plate storage position for storing multi-well plates required in the gene sequencing sample preparation process; a product storage position for storing product plates loaded with the product samples; and a consumable stack position for storing a certain number of boxed tips and / or plate consumables.

[0007] In some possible embodiments, the main housing includes a shell and an operating table located inside the shell, and along the length direction of the operating table, the operating table is divided into a first area, a second area, a third area, a fourth area, a fifth area and a sixth area arranged side by side, wherein the sample storage position is arranged in the first area, the tip storage position is arranged in the second area, the plate storage position and part of the reagent storage position are arranged in the third area, another part of the reagent storage position is arranged in the fourth area, the carrier plate storage position and the product storage position are arranged in the fifth area, and the consumables stacking position is arranged in the sixth area.

[0008] In some possible embodiments, the droplet generation module is arranged in the sixth area, located at the end of the consumables stacking position and adjacent to the carrier storage position, the quantification module is located in the third area, the biochemical reaction module includes a PCR instrument, a purification and sorting device, and a temperature-controlled oscillation device, the PCR instrument is located in the fourth area near the end of the droplet generation module and adjacent to the quantification module, the purification and sorting device is located in the third area, the temperature-controlled oscillation device is located in the fourth area and at the end away from the PCR instrument, and the purification and sorting device and the temperature-controlled oscillation device are adjacent to the tip storage position and the reagent storage position.

[0009] In some possible embodiments, the main housing further includes an automatic window and door assembly provided on one side of the shell, and the automatic window and door assembly is used to seal the internal cavity of the shell when closed.

[0010] In some possible embodiments, the scheduling and transfer module includes a robotic arm device disposed in the main housing, and a gripping device and multiple independent pipetting devices disposed on the robotic arm device. The robotic arm device can drive the gripping device and each of the pipetting devices to move along a first direction and a second direction perpendicular to each other.

[0011] In some possible embodiments, the grasping device includes: a grasping mounting plate provided on the robotic arm device, a grasping drive assembly provided on the grasping mounting plate, and a grasping assembly provided on the grasping drive assembly, wherein the grasping drive assembly can drive the grasping assembly to move along a third direction, wherein the third direction is perpendicular to a plane formed by the first direction and the second direction, and the grasping drive assembly can also drive the grasping assembly to rotate within the plane.

[0012] In some possible embodiments, a pipetting mounting plate is provided on the robotic arm device, and a plurality of the pipetting devices are independently arranged on the pipetting mounting plate, and each of the pipetting devices includes a pipetting drive component, a pipetting control circuit board electrically connected to the pipetting drive component, and a pipette provided on the pipetting drive component.

[0013] In some possible embodiments, the automated sample preparation system further includes a tabletop code scanning device disposed near the consumables storage module, and a robotic arm code scanning device disposed on the scheduling and transfer module.

[0014] In some possible embodiments, the automated sample preparation system further includes a control module, which is configured to control the coordinated operations of the consumables storage module, the droplet generation module, the biochemical reaction module, the quantification module, and the scheduling and transfer module.

[0015] In summary, the automated sample preparation system provided in the embodiments of the present application implements a one-click, automated single-cell sample preparation process. After the sample is input, a library sample for gene sequencing can be output, reducing human intervention, minimizing manual operation errors, saving manpower, and lowering costs. Furthermore, each module can perform pipeline operations in parallel, improving the throughput and efficiency of library construction. The entire process is carried out in a sealed chamber, avoiding environmental pollution and cross-contamination between samples, improving the quality of prepared gene sequencing samples, and making the results more accurate and reliable. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a system framework diagram of an automated sample preparation system provided in one embodiment of the present application.

[0017] Figure 2 This is a schematic diagram of the structure of an automated sample preparation system provided in one embodiment of the present application.

[0018] Figure 3 for Figure 2 Schematic diagram of the structure of the main chassis.

[0019] Figure 4 yes Figure 3 Schematic diagram of the structure after the transparent window door is closed.

[0020] Figure 5 yes Figure 3 Schematic diagram of the structure after the transparent window door is opened.

[0021] Figure 6 yes Figure 2 Schematic diagram of the three-dimensional structure of the operating table and each module.

[0022] Figure 7 for Figure 2 Top view of the operating table and each module.

[0023] Figure 8 for Figure 7 Schematic diagram of consumables placed on each consumable storage location.

[0024] Figure 9 for Figure 2 Schematic diagram of the structure of the scheduling transfer module.

[0025] Figure 10 for Figure 9 Schematic diagram of the structure of the grabbing device.

[0026] Figure 11 for Figure 9 Schematic diagram of the structure of the pipetting device.

[0027] Figure 12 for Figure 3 Schematic diagram of the structure of the top electronic control component.

[0028] Figure 13 for Figure 3 Schematic diagram of the structure of the mid-bottom electronic control component.

[0029] Figure 14 A schematic diagram of the process of preparing a single cell sample according to an embodiment of the present application.

[0030] Description of main component symbols

[0031] Automated sample preparation system 100 Second moving component 612

[0032] Main housing 1 bracket 613

[0033] Housing 11 Mounting frame 614

[0034] Window 111 Grab drive assembly 615

[0035] Operation table 12 pipetting drive assembly 616

[0036] First zone 121 grabbing device 62

[0037] Second zone 122 grabs the mounting plate 621

[0038] Third zone 123 grab drive assembly 622

[0039] Fourth zone 124 grabbing component 623

[0040] Area 5 125 Robotic Arm Scanning Device 624

[0041] Sixth area 126 pipetting device 63

[0042] Automatic window and door assembly 13 Pipetting drive assembly 631

[0043] Transparent window door 131 pipetting control circuit board 632

[0044] Window and door detection sensor 132 Pipette 633

[0045] Adsorption parts 133 Pipetting mounting bracket 64

[0046] First window 134 control module 7

[0047] Second window door 135 top electric control component 71

[0048] Connector 136 Computer host 711

[0049] Strengthen door frame 137 Switching power supply 712

[0050] UV sterilizer lamp 14 control board 713

[0051] Air filter 15 relay board 714

[0052] Consumables storage module 2 Ballast 715

[0053] Sample storage position 21 bottom electronic control component 72

[0054] Reagent storage location 22 PCR controller 721

[0055] Normal temperature reagent storage position 221 solid state relay 722

[0056] Low temperature reagent storage position 222 Power switch 723

[0057] Reagent slot 223 Temperature control board 724

[0058] Tip storage position 23 Human-computer interaction module 8

[0059] Carrier storage position 24 support platform 81

[0060] Plate storage position 25 Display 82

[0061] Microplate position 251 Tabletop code scanning device 9

[0062] Deep well plate position 252 Sample container 101

[0063] Consumables stack 26 reagent containers 102

[0064] Product storage position 27 Tip 103

[0065] Sealing cover temporary storage position 28 Chip carrier 104

[0066] Waste storage 29 Microplate 105

[0067] Droplet generation module 3 deep well plate 106

[0068] Biochemical reaction module 4 product plate 107

[0069] PCR instrument 41 PCR plate 108

[0070] Purification and sorting device 42 PCR pad 109

[0071] Temperature control oscillation device 43 first direction X

[0072] Quantitative module 5 Second direction Y

[0073] Scheduling transfer module 6 Third direction Z

[0074] The length direction a of the robot arm device 61

[0075] First moving component 611 width direction b

[0076] The following specific implementation methods will further illustrate the present application in conjunction with the above-mentioned drawings. DETAILED DESCRIPTION

[0077] The following will clearly and completely describe the technical solutions in the embodiments of the present application in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments.

[0078] It should be noted that when a component is referred to as being "fixed to" or "mounted on" another component, it may be directly on the other component or there may be a central component. When a component is referred to as being "disposed on" another component, it may be directly on the other component or there may be a central component. As used herein, the term "and / or" includes all and any combinations of one or more of the relevant listed items.

[0079] It should be noted that, although a logical order is shown in the flowchart, in some cases, the steps shown or described may be performed in an order different from that in the flowchart. The method disclosed in the embodiments of the present application includes one or more steps or actions for implementing the method. The method steps and / or actions may be interchangeable with each other without departing from the scope of the claims. Unless a specific order of steps or actions is specified, the order and / or use of specific steps and / or actions may be modified without departing from the scope of the claims.

[0080] See also Figure 1 and Figure 2 Figure 1 shows a schematic diagram of an automated sample preparation system 100 in one embodiment of the present application. Automated sample preparation system 100 is used for sample preparation during gene sequencing, specifically, for constructing sequencing libraries. Automated sample preparation system 100 includes a consumables storage module 2, a droplet generation module 3, a biochemical reaction module 4, a quantification module 5, and a scheduling and transfer module 6.

[0081] Consumables storage module 2 is used to store consumables (see Figure 8). Consumables can be used to load biological samples (such as single cell samples: cell (nucleus) / protoplast suspension), and can also be used to load various connectors and reagents required for gene sequencing sample preparation. They can also be used to be installed in sample carriers (such as magnetic beads) for carrying biological samples in sequencing library construction, but are not limited to this. The consumables can be, but are not limited to, tube consumables (such as sample tubes, reagent tubes, centrifuge tubes, etc.), pipette tips 103, multi-well plates (such as microplates 105, deep-well plates 106, etc.), product plates 107, chip carriers 104, PCR plates 108, PCR pads 109 used by PCR instruments, etc. The droplet generation module 3 can be used to prepare single cell suspensions into microdroplets. The biochemical reaction module 4 includes a plurality of biochemical reaction devices. The entire process of gene sequencing sample preparation is divided into a plurality of different sub-processes, and each biochemical reaction device is used to execute one or more of the sub-processes. For example, multiple biochemical reaction devices are used to execute different sub-processes, that is, multiple biochemical reaction devices are in a one-to-one correspondence with multiple sub-processes, or a certain biochemical reaction device can be set to correspond to at least two sub-processes, or a certain sub-process can be set to correspond to at least two biochemical reaction devices. The quantification module 5 can quantify the product samples in the biochemical reaction process, where the product samples can be intermediate products after a certain sub-process or final products. The scheduling and transfer module 6 can move between the consumables storage module 2, the droplet generation module 3, the biochemical reaction module 4, and the quantification module 5 to transfer the consumables 10, so that the droplet generation module 3 completes the micro-droplet preparation, and the multiple biochemical reaction devices successively complete the corresponding biochemical reaction sub-processes, and complete the quantification of the intermediate products and final products of the biochemical reaction process, thereby completing the preparation of automated gene sequencing samples. The automated sample preparation system 100 also includes a control module 7 for controlling the coordinated operations of the consumables storage module 2, the droplet generation module 3, the biochemical reaction module 4, the quantification module 5, and the scheduling and transfer module 6.

[0082] For single-cell samples, before gene sequencing, the first step is to prepare microdroplets and reverse transcription within the droplets; the second step is to break the emulsion, enrich and purify, sort the oligo and cDNA, and use the polymerase chain reaction (PCR) to amplify the cDNA, purify, and quantify; the third step is to PCR amplify the oligo products, fragment screening, cDNA shearing, end repair, ligation of molecular tag adapters, purification, fragment screening, PCR amplification of the cDNA products, purification, and quantification; the fourth step is to perform rolling circle amplification of the library to obtain nanospheres (DNBs), quantification, and other processes, and finally, a library for gene sequencing can be obtained. Therefore, in this embodiment, the droplet generation module 3 is used to perform the microdroplet preparation process in the first step, the quantification module 5 is used to perform the quantification process in each step, and multiple biochemical reaction devices are used to perform other sub-processes in the first to fourth steps except droplet generation and quantification. Depending on the type of sample, the above-mentioned gene sequencing sample preparation process can also be modified.

[0083] like Figures 1 to 3 As shown, the automated sample preparation system 100 includes a main housing 1, in which a consumables storage module 2, a droplet generation module 3, a biochemical reaction module 4, a quantitative module 5, and a scheduling and transfer module 6 are integrated. Specifically, the main housing 1 includes a shell 11 and an operating table 12 located within the shell 11. The aforementioned multiple functional modules are integrated on the operating table 12, so that the automated sample preparation process can be carried out in the inner cavity of the shell 11. Figures 2 to 5 As shown, the main housing 1 also includes an automatic window and door assembly 13 provided on one side of the shell 11. The automatic window and door assembly 13 is used to seal the internal cavity of the shell 11 when closed, so as to isolate the sample preparation process from the external environment. In some embodiments, the automatic window and door assembly 13 includes a transparent window door 131, a window and door detection sensor 132, and an adsorbent 133. The transparent window door 131 is provided at the window 111 on one side of the shell 11, which can facilitate the operator to observe the movement of the scheduling transfer module 6 in the shell 11 and the progress of sample preparation. The window and door detection sensor 132 is provided at the window 111, which can detect whether the transparent window door 131 is open or closed. The adsorbent 133 is provided at the bottom of the window 111, which can provide magnetic force to hold the transparent window door 131 when it is closed, preventing the transparent window door 131 from being easily opened.

[0084] In some embodiments, the transparent window door 131 can be flipped open and closed. Figure 4 As shown, the transparent window door 131 is closed. Figure 5 As shown, the transparent window door 131 is flipped open. This flip structure is easy to operate and saves space.

[0085] In some embodiments, the transparent window door 131 includes a first window door 134, a second window door 135, a connector 136, a reinforced door frame 137, and a handle 138. The first window door 134 and the second window door 135 are connected by a plurality of connectors 136. The handle 138 is located on the edge of the second window door 135. The first window door 134 is mounted on a reinforced door frame 137, which is mounted on the window 111 of the housing 11. The flip design of the two windows facilitates opening and closing, saves operating space, and enhances the appearance.

[0086] An ultraviolet sterilization lamp 14 may also be provided in the housing 11 , and the ultraviolet sterilization lamp 14 is used to perform ultraviolet sterilization and disinfection on the internal cavity of the housing 11 .

[0087] An air filter 15 is provided on the top of the housing 11 to filter the air entering the housing 11 and provide positive pressure inside the housing 11 to prevent cross contamination between the sample and the environment. Specifically, the air filter 15 can be a laminar flow hood.

[0088] like Figures 6 to 8 As shown, the consumable storage module 2 includes multiple consumable storage locations, specifically including a sample storage location 21, a reagent storage location 22, a pipette tip storage location 23, a carrier plate storage location 24, a plate storage location 25, a product storage location 27, and a consumable stack location 26. The sample storage location 21 is used to store a sample container 101 containing a biological sample. For example, the sample container 101 can be a sample tube, and the corresponding sample storage location 21 can be a sample tube rack for placing the sample tube. The reagent storage location 22 is used to store a reagent container 102 containing reagents required for a biochemical reaction. For example, the reagent container 102 can be a reagent tube or a reagent tank, and the corresponding reagent storage location 22 can be a reagent tube rack for placing the reagent tube, or a reagent tank rack for placing the reagent tank. The pipette tip storage location 23 is used to store pipette tips 103 for transferring the biological sample or the reagent. For example, the pipette tip storage location 23 can be a pipette tip rack. The carrier plate storage location 24 is used to store the chip carrier plate 104 required for droplet generation. Plate storage 25 is used to store the multiwell plates required for gene sequencing sample preparation. Multiwell plates can include conventional microplates 105 and deep-well plates 106. Product storage 27 is used to store product plates 107 loaded with products. Consumables stacking 26 is used to store a certain number of boxed pipette tips 103 and at least one of some plate-type consumables (such as PCR plates 108, PCR pads 109, and deep-well plates 106). In addition, consumables stacking 26 is also equipped with an empty transfer position and a waste plate position for stacking discarded plate-type consumables.

[0089] In some embodiments, the consumables storage module 2 further includes a temporary sealing cap storage location 28 and a waste storage location 29. The temporary sealing cap storage location 28 is used to temporarily store sealing caps removed from tube-type consumables, while the waste storage location 29 is used to collect discarded consumables and waste liquids.

[0090] The automated sample preparation process utilizes a large number of consumables, making their storage on the work surface 12 a key design consideration. To facilitate the handling and dispatch of consumables, while also improving space utilization and minimizing the overall size of the machine, in this embodiment, the consumables are strategically arranged in corresponding areas of the work surface 12 according to the sequence of biochemical reaction processes. The work surface 12 is divided into multiple functional areas, specifically a first area 121, a second area 122, a third area 123, a fourth area 124, a fifth area 125, and a sixth area 126, arranged in parallel along the length direction a of the work surface 12. The sample storage area 21 is located at one end of the operating table 12 (i.e., the first area 121). Multiple tip storage areas 23 can be provided, with multiple tip storage areas 203 arranged side by side in the second area 122. The plate storage area 25 and some reagent storage areas 22 are located in the third area 123, while another portion of the reagent storage areas 22 are located in the fourth area 124. The carrier plate storage area 24 and product storage area 27 are located in the fifth area 125. A consumables stacking area 26 can be located at the other end of the operating table 12 (i.e., the sixth area 126). Furthermore, a temporary sealing cap storage area 28 and a waste storage area 29 are both located in the fifth area 125.

[0091] In some embodiments, the sample storage location 21 is configured in an elongated strip shape, capable of accommodating a row of sample containers 101 at one end of the operating table 12. For example, the sample storage location 21 may be a long, rectangular sample rack having a row of through-holes for accommodating sample tubes. Specifically, in this embodiment, the automated sample preparation system 100 can achieve automated library construction for 1-16 samples, with the sample storage location 21 having at least 16 through-holes for accommodating sample tubes.

[0092] In some embodiments, the tip storage position 23 is a roughly rectangular porous mounting rack, and multiple rectangular porous mounting racks are arranged side by side on one side of the long strip sample storage position 21. Tips are the most frequently used consumables in the sample preparation process and are the largest in number. Therefore, the entire second area 122 is used to set up the tip storage position 23, and the tip storage position 23 is set between the sample storage position 21 and the reagent storage position 22 to facilitate the use of tips, shorten the distance for taking and placing tips, and improve efficiency.

[0093] In some embodiments, depending on the type of reagents stored, the reagent storage area 22 primarily includes a room-temperature reagent storage area 221, a low-temperature reagent storage area 222, and a reagent slot 223. Specifically, the reagent slot 223 is located in the third area 123 adjacent to the tip storage area 23, while the room-temperature reagent storage area 221 and the low-temperature reagent storage area 222 are located side by side in the fourth area 124.

[0094] In some embodiments, depending on the type of plates stored, the plate storage position 25 mainly includes a microplate position 251 for placing an ordinary microplate 105 and a deep-well plate position 252 for placing a deep-well plate 106. The ordinary microplate position 251 and the deep-well plate position 252 are arranged side by side with the reagent slot 233 in the third area 123.

[0095] In some embodiments, the operating table 12 further includes identification marks (not shown) corresponding to each area, which are used to distinguish different consumables and effectively avoid incorrect use of consumables.

[0096] like Figures 6 to 8 As shown, the droplet generation module 3 can specifically be a microfluidic droplet generation device. Based on the microfluidic droplet generation principle, the chip carrier 104 can be used to prepare single-cell suspensions into microdroplets on the microfluidic droplet generation device. Specifically, the microfluidic droplet generation device can simultaneously generate droplets for multiple samples, for example, eight samples. If 16 samples need to be processed simultaneously, two rounds of droplet generation are required.

[0097] The structures of the multiple biochemical reaction devices in the biochemical reaction module 4 are different according to the different sub-processes to be executed. The following is an illustration of the construction of a single-cell library. The multiple biochemical reaction devices may specifically include: a PCR instrument 41, a purification and sorting device 42, and a temperature-controlled oscillation device 43. Among them, the PCR instrument 41 can be used to execute reaction processes such as RT reaction and PCR amplification. The purification and sorting device 42 can be used to execute processes such as enrichment, purification, and sorting. The temperature-controlled oscillation device 43 can be used to execute an oscillation incubation process, and "incubation" is to create the required temperature conditions for the biochemical reaction of the biological sample by heating, cooling, and keeping warm. Specifically, the purification and sorting device 42 and the temperature-controlled oscillation device 43 of the embodiment of the present application can simultaneously execute the corresponding sub-process operations of multiple samples, for example, the corresponding sub-process operations of 1-16 samples can be executed.

[0098] The quantitative module 5 can be a quantitative detection device, such as a fluorescence quantitative detection device. The quantitative module 5 is combined with the scheduling transfer module 6 to transfer the intermediate products and final products in the single-cell library construction process to the quantitative module 5 for quantitative detection, without interrupting the sample preparation process or manually removing the intermediate products for quantitative detection, thereby realizing full-process automated sample preparation.

[0099] Based on the aforementioned layout of the various storage locations in the consumables storage module 2, the droplet generation module 3 is positioned in the sixth zone 126, at the end of the consumables stacking location 26 and adjacent to the carrier storage location 24. That is, along the width direction b of the operating table 12, the droplet generation module 3 and the consumables stacking location 26 are arranged sequentially. The PCR instrument 41 is located at the end of the fourth zone 124 near the droplet generation module 3, and the temperature-controlled oscillator 43 is located in the fourth zone 124, at the end away from the PCR instrument 41. That is, along the width direction b, the PCR instrument 41, the low-temperature reagent storage location 222, the room-temperature reagent storage location 221, and the temperature-controlled oscillator 43 are arranged sequentially in the fourth zone 124. The purification and sorting device 42 and the quantification module 5 are both located in the third zone 123. Furthermore, the purification and sorting device 42 and the temperature-controlled oscillating device 43 are positioned adjacent to the tip storage position 23 and the reagent storage position 22. Specifically, the purification and sorting device 42 is located between the microplate position 251 and the reagent slot 223, and the quantification module 5 is located between the microplate position 251 and the deep-well plate position 252. The quantification module 5 is positioned adjacent to the PCR instrument 41. By rationally arranging the various modules on the operating table 12, the travel distance of the dispatching and transfer module 6 is effectively shortened, optimizing the preset dispatching and transfer paths, thereby improving the operational convenience and efficiency of the sample preparation process. Furthermore, the rational and scientific layout of the various modules effectively increases the space utilization of the operating table 12, facilitating the miniaturization of the automated sample preparation system 100.

[0100] like Figures 9 to 11 As shown, combined Figure 6 The scheduling and transfer module 6 includes a robotic arm device 61, a gripping device 62 and a plurality of independent pipetting devices 63 arranged on the robotic arm device 61. The robotic arm device 61 can drive the gripping device 62 and each pipetting device 63 to move along a first direction X and a second direction Y perpendicular to each other. Among them, the robotic arm device 61 is on the operating table 12, the first direction X can be the length direction a of the operating table 12, and the second direction Y can be the width direction b of the operating table 12. The gripping device 62 can move along a third direction Z, and the third direction Z is perpendicular to the plane formed by the first direction X and the second direction Y, and the gripping device 62 can also rotate within the plane formed by the first direction X and the second direction Y to grab or release consumables. The pipetting device 63 can move along the third direction Z and realize liquid absorption and discharge, and can realize automatic transfer of liquid in conjunction with the robotic arm device 61.

[0101] like Figure 6 and Figure 9As shown, the robotic arm device 61 includes a first movable assembly 611 disposed on the operating table 12 and a second movable assembly 612 disposed on the first movable assembly 611. The gripping device 62 and multiple independent pipetting devices 63 are all disposed on the second movable assembly 612. The first movable assembly 611 is disposed at the edge of the operating table 12 and is capable of moving along a first direction X. The second movable assembly 612 specifically includes a bracket 613 disposed on the first movable assembly 611, a mounting frame 614 disposed on the bracket 613, a gripping drive assembly 615 disposed on the mounting frame 614, and a pipetting drive assembly 616 disposed on the mounting frame 614. The gripping device 62 is disposed on the gripping drive assembly 615, which is capable of driving the gripping device 62 to move along a second direction Y. The multiple pipetting devices 63 are integrally mounted on the pipetting drive assembly 616 via the pipetting mounting frame 64. The pipetting drive assembly 616 is capable of driving the multiple pipetting devices 63 to move synchronously along the second direction Y.

[0102] In some embodiments, the grabbing drive assembly 615 and the pipetting drive assembly 616 are arranged side by side along the first direction X, wherein the grabbing drive assembly 615 is located on the outside of the mounting frame 614 and the pipetting drive assembly 616 is located on the inside of the mounting frame 614 .

[0103] like Figure 9 and Figure 10 As shown, the gripping device 62 includes a gripping mounting plate 621 mounted on the robotic arm assembly 61, a gripping drive assembly 622 mounted on the gripping mounting plate 621, and a gripping assembly 623 mounted on the gripping drive assembly 622. Specifically, the gripping mounting plate 621 is mounted on the gripping drive assembly 615. The gripping drive assembly 622 can drive the gripping assembly 623 to move along the third direction Z and can also drive the gripping assembly 623 to rotate within a plane defined by the first direction X and the second direction Y. Furthermore, the gripping device 62 can also open and close the sealing cover on the container through rotational motion.

[0104] In some embodiments, the gripping device 62 also includes a robotic arm code scanning device 624 disposed on the gripping mounting plate 621 and corresponding to the gripping component 623, which can scan one-dimensional or two-dimensional codes on plate position consumables such as the plate storage position 25 and the carrier storage position 24 on the operating table 12.

[0105] like Figure 9 and Figure 11As shown, the pipetting device 63 includes a pipetting drive assembly 631, a pipetting control circuit board 632 electrically connected to the pipetting drive assembly 631, and a pipette 633 disposed on the pipetting drive assembly 631. The pipetting drive assembly 631 can drive the pipette 633 to move up and down along a third direction Z, thereby extending the pipette 633 into or out of a container containing liquid. The pipetting control circuit board 632 can control the pipette 633 to absorb or discharge liquid to complete liquid transfer. Multiple pipetting devices 63 can achieve independent movement along the third direction Z, thereby enabling each pipetting device 63 to independently extract or discharge liquid.

[0106] like Figure 12 and Figure 13 As shown, combined Figures 1 to 3 The control module 7 includes a top electric control component 71 and a bottom electric control component 72 , wherein the top electric control component 71 is arranged at the top of the shell 11 , and the bottom electric control component 72 is arranged at the bottom of the shell 11 and is located below the operating table 12 .

[0107] In some embodiments, the top electronic control assembly 71 includes a computer host 711, a switching power supply 712, a control board 713, a relay board 714, and a ballast 715, all of which are mounted on the top plate of the housing 11. The computer host 711 is the main computer that runs the entire device and primarily controls its operation. The switching power supply 712 provides power to the electronic components. The control board 713 provides hardware control programs for the electronic components.

[0108] In some embodiments, the bottom electronic control assembly 72 includes a PCR controller 721, a solid-state relay 722, a power switch 723, and a temperature control board 724. The PCR controller 721 is used to control the PCR instrument 41. The temperature control board 724 is used to control the low-temperature reagent storage location 222 and the temperature-controlled oscillating device 43.

[0109] like Figures 6 to 8 As shown, the automated sample preparation system 100 also includes a tabletop barcode scanning device 9, which is arranged near the sample storage position 21. The tabletop barcode scanning device 9 is used to scan the position barcode of each well position on the sample storage position 21 and the one-dimensional code on the tube consumables placed at the well position.

[0110] like Figure 2 As shown, the automated sample preparation system 100 may further include a human-computer interaction module 8. The human-computer interaction module 8 includes a support platform 81 disposed on one side of the housing 11 and a display 82 located on the support platform 81. The display 82 is used for an operator to monitor the progress of the biochemical reaction in the housing 11, so that manual intervention can be carried out in a timely manner when an abnormality occurs.

[0111] The automated sample preparation system 100 of the embodiment of the present application is further described below using the 3' RNA single cell library construction process as an example. The automated sample preparation system 100 can automatically construct libraries for 16 samples.

[0112] First, place the sample container 101 (specifically, sample tubes, 1-16) loaded with biological samples in the corresponding holes of the sample storage position 21, and place other reagents in the room temperature reagent storage position 221, the low temperature reagent storage position 222 and the reagent slot 223.

[0113] At the beginning of the process, select the process type 3'RNA and the process node (3'RNA is divided into 4 sub-steps, such as Figure 14 As shown), follow the UI prompts (operating panel 12 to Figure 8 For example, place the corresponding number of reagents and consumables, and scan the biological samples and product plates.

[0114] Step 1, Droplet Generation and In-Droplet Reverse Transcription: The robotic arm 61 drives the pipetting device 63 to transfer the biological sample, magnetic bead phase, and droplet generation oil to the chip carrier 104. The robotic arm 61 then drives the gripping device 62 to place the chip carrier 104 into the droplet generation module 3 to complete microdroplet generation (if more than eight samples are needed, two rounds of microdroplet generation are performed). After microdroplet generation, the robotic arm 61 drives the pipetting device 63 to transfer the microdroplets to the PCR instrument 41 for in-droplet reverse transcription (RT). Once the RT reaction is complete, Step 1 ends.

[0115] Step 2, emulsion breaking and cDNA amplification: After the RT reaction is completed, the pipetting device 63 is driven by the robotic arm device 61 to transfer the microdroplets to the deep well plate 106, and then PFO is added to break the emulsion of the microdroplets. After emulsion breaking, magnetic beads are added to enrich Oligo (sequences on small magnetic beads used to distinguish cells) and cDNA (DNA complementary to RNA after reverse transcription), and Oligo and cDNA are sorted in the purification and sorting device 42 by adding different volumes of purification magnetic beads. Oligo is purified to obtain Oligo products and placed on the product plate 107. After cDNA purification, it needs to be transferred to the PCR instrument 41 for amplification. The amplified cDNA is then purified in the purification and sorting device 42 to obtain cDNA products, and placed on another product plate 107. Finally, the two products are quantified in the quantitative module 5, and step 2 ends here.

[0116] Step 3, respectively, build libraries for the Oligo product and cDNA product: The robotic arm device 61 drives the pipetting device 63 to absorb the Oligo product and transfer it to the PCR chip, add the reagents required for amplification to perform a PCR reaction, and at the same time prepare the cDNA shearing reagent mixture. After the Oligo library is built, fragment screening is performed in the purification and sorting device 42, and then the Oligo library is obtained and placed on a new product plate 107. After cDNA shearing, end repair and linker connection are required, and then the connection product is purified and fragment screening is performed. After the chip screening, PCR amplification is performed, followed by purification to obtain the cDNA library. Finally, the cDNA library is quantified, and step 3 is completed.

[0117] In step 4, the Oligo library and cDNA library are mixed (Pooling) and replicated into DNA nanoballs (DNBs) through a one-step rolling circle method. The DNBs are quantified and then the DNBs can be sequenced.

[0118] One or more of the above four steps can be selected for execution, but the selected steps must be continuous.

[0119] In addition, the automated sample preparation system 100 of the present application can also realize the construction of libraries for single-cell 5'RNA, ATAC and other omics. Therefore, the system can realize the whole process from cell suspension (cell, nucleus, protoplast) to DNB in ​​one click, and is compatible with omics such as single-cell 3'RNA, 5'RNA, ATAC, and can complete 1-16 flexible samples, each sample containing tens of thousands of cells (nucleus, protoplast), so as to analyze the heterogeneity of similar cells. Truly realize the automation of the whole process from prepared single-cell (nucleus, protoplast) suspension to pre-sequencing library and DNB preparation.

[0120] In summary, the automated sample preparation system 100 provided in the embodiments of the present application implements a one-click, automated single-cell sample preparation process. After the sample is input, a library sample for genetic sequencing can be output, reducing human intervention, minimizing manual operation errors, saving manpower, and lowering costs. In particular, the coordination of the quantification module 5 and the scheduling and transfer module 6 enables concentration detection of intermediate and final products. Furthermore, each module can perform pipeline operations in parallel, improving the throughput and efficiency of library construction. Furthermore, the entire process is carried out within a sealed chamber, avoiding environmental contamination and cross-contamination between samples, improving the quality of prepared genetic sequencing samples, and making the results more accurate and reliable.

[0121] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application and are not intended to limit the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present application.

Claims

1. An automated sample preparation system, characterized in that: include: A main housing and a consumable storage module, a droplet generation module, a biochemical reaction module, a quantitative module and a scheduling and transfer module integrated in the main housing, Wherein, the consumables storage module is configured to store consumables; The droplet generation module is configured to generate microdroplets containing biological samples; The biochemical reaction module includes a plurality of biochemical reaction devices, each of which is used to execute one or more sub-processes in the gene sequencing sample preparation process; The quantitative module is configured to quantitatively detect the concentration of the product sample obtained during the gene sequencing sample preparation process; The scheduling and transfer module is configured to grab the consumables stored in the consumable storage module, and the scheduling and transfer module is also configured to move along a predetermined path between the consumable storage module, the droplet generation module, the biochemical reaction module and the quantitative module to complete the transfer of the consumables and / or liquids.

2. The automated sample preparation system according to claim 1, wherein: The consumables storage module includes multiple consumables storage locations for storing multiple consumables, and the multiple consumables storage locations include: A sample storage location, used for storing a sample container loaded with a biological sample; Reagent storage location, used to store reagent containers loaded with reagents required for biochemical reactions; a pipette tip storage position, used for storing pipette tips for transferring the biological sample or the reagent; A carrier storage location for storing the chip carrier required for droplet generation; Plate storage location, used to store the multi-well plates required in the gene sequencing sample preparation process; a product storage location, for storing a product plate loaded with the product sample; and Consumables stacking position, used to store a certain number of boxed tips and / or plate-type consumables.

3. The automated sample preparation system according to claim 2, wherein: The main housing includes a shell and an operating table located inside the shell. Along the length direction of the operating table, the operating table is divided into a first area, a second area, a third area, a fourth area, a fifth area and a sixth area arranged side by side. The sample storage position is arranged in the first area, the tip storage position is arranged in the second area, the plate storage position and part of the reagent storage position are arranged in the third area, another part of the reagent storage position is arranged in the fourth area, the carrier plate storage position and the product storage position are arranged in the fifth area, and the consumables stacking position is arranged in the sixth area.

4. The automated sample preparation system according to claim 3, wherein: The droplet generation module is arranged in the sixth area, located at the end of the consumables stacking position and adjacent to the carrier storage position. The quantification module is located in the third area. The biochemical reaction module includes a PCR instrument, a purification and sorting device, and a temperature-controlled oscillation device. The PCR instrument is located in the fourth area near the end of the droplet generation module and adjacent to the quantification module. The purification and sorting device is located in the third area. The temperature-controlled oscillation device is located in the fourth area and at the end away from the PCR instrument. The purification and sorting device and the temperature-controlled oscillation device are adjacent to the pipette tip storage position and the reagent storage position.

5. The automated sample preparation system according to claim 3, wherein: The main housing further comprises an automatic window and door assembly provided on one side of the shell, and the automatic window and door assembly is used to seal the internal cavity of the shell when closed.

6. The automated sample preparation system according to claim 1, wherein: The scheduling and transfer module includes a robotic arm device arranged in the main housing, a grasping device and multiple independent pipetting devices arranged on the robotic arm device, and the robotic arm device can drive the grasping device and each of the pipetting devices to move along a first direction and a second direction perpendicular to each other.

7. The automated sample preparation system according to claim 6, wherein: The grasping device includes: a grasping mounting plate arranged on the robotic arm device, a grasping drive assembly arranged on the grasping mounting plate, and a grasping assembly arranged on the grasping drive assembly. The grasping drive assembly can drive the grasping assembly to move along a third direction, and the third direction is perpendicular to the plane formed by the first direction and the second direction. The grasping drive assembly can also drive the grasping assembly to rotate within the plane.

8. The automated sample preparation system according to claim 6, wherein: A pipetting mounting plate is provided on the robotic arm device, and multiple pipetting devices are independently arranged on the pipetting mounting plate. Each pipetting device includes a pipetting drive component, a pipetting control circuit board electrically connected to the pipetting drive component, and a pipette provided on the pipetting drive component.

9. The automated sample preparation system according to claim 1, wherein: The automated sample preparation system further includes a tabletop code scanning device disposed near the consumables storage module, and a robotic arm code scanning device disposed on the scheduling and transfer module.

10. The automated sample preparation system according to claim 1, wherein: The automated sample preparation system further includes a control module, which is configured to control the consumables storage module, the droplet generation module, the biochemical reaction module, the quantification module, and the scheduling and transfer module to coordinate operations.

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