An in situ transcriptomic-based tissue sequencing sample automated processing apparatus

Abstract

The application relates to the field of in-situ fluorescent sequencing technology equipment, in particular to an automatic processing equipment for tissue sequencing samples based on in-situ transcription technology. The automatic processing equipment comprises a communication device, the communication device comprises a syringe pump, a pump body butt joint, a rotating butt joint mechanism and a lifting mechanism; the lifting mechanism drives the pump body butt joint and the rotating butt joint mechanism to move up and down synchronously; the rotating butt joint mechanism comprises a clamping joint and a first motor, and the first motor drives the clamping joint to rotate; a reagent disc device comprises a reagent disc, a mounting seat, a chip connector and a gating valve, the reagent disc is provided with a plurality of liquid storage pools, and the chip connector is mounted on the mounting seat; the gating valve can rotate to select and open one of the liquid storage pools to be communicated with the chip connector; the mounting seat is provided with a pump body connecting hole, the pump body connecting hole is communicated with the chip connector; and the lifting mechanism can drive the pump body butt joint to be butt jointed with the pump body connecting hole and the clamping joint to be butt jointed with the gating valve. The application has the advantages of improving the preparation efficiency of sequencing samples.

Description

Technical Field

[0001] This application relates to the field of fluorescence in situ sequencing technology equipment, and in particular to an automated tissue sequencing sample processing device based on in situ transcriptomics. Background Technology

[0002] Spatial transcriptomics can locate and differentiate the active expression of functional genes in specific tissue regions, thus providing important information for basic research and clinical diagnosis. As a groundbreaking new omics research technology, it enables us to detect gene activity in different microenvironments within tissue samples and to map the spatial expression of active genes.

[0003] Current spatial transcriptomics methods are mainly divided into two categories: sequencing-based methods and microscopic imaging-based methods. Microscopic imaging-based methods perform in situ sequencing or multiple rounds of single-molecule fluorescence in situ hybridization and imaging at the original location of RNA in cells or tissues, thereby obtaining a series of signals encoded by different fluorescent colors to detect different genes and directly obtain the spatial location information of the detected genes.

[0004] In situ sequencing based on the aforementioned spatial transcriptomics requires incubation and staining of tissue cell sections to prepare sequencing samples. However, current methods typically involve manually adding various reagents directly to a glass slide containing the tissue sample for sequencing sample preparation, resulting in low efficiency. Summary of the Invention

[0005] To improve the efficiency of sequencing sample preparation, this application provides an automated tissue sequencing sample processing device based on in situ transcriptomics.

[0006] The automated tissue sequencing sample processing device based on in situ transcriptomics provided in this application adopts the following technical solution: an automated tissue sequencing sample processing device based on in situ transcriptomics, comprising: a support frame;

[0007] A connecting device includes an injection pump, a pump body connector, a rotary docking mechanism, and a lifting mechanism. The injection pump and the lifting mechanism are mounted on a support frame. The pump body connector is connected to the injection pump via a pipeline. The lifting mechanism drives the pump body connector and the rotary docking mechanism to move up and down synchronously. The rotary docking mechanism includes a snap-fit ​​connector and a first motor, and the first motor drives the snap-fit ​​connector to rotate.

[0008] A reagent tray device includes a reagent tray, a mounting base, a chip connector, and a gate valve. The mounting base is fixedly disposed on one side of the reagent tray, and the reagent tray has multiple liquid reservoirs spaced apart along its circumference. The chip connector has a reaction chamber for incubating tissue samples and is detachably mounted on the mounting base. The gate valve is disposed on the reagent tray and can rotate to select one of the liquid reservoirs to communicate with the chip connector. The mounting base is provided with a pump connection hole, which communicates with the chip connector.

[0009] The reagent tray is located on the lower side of the support frame. The reagent tray device is placed on the reagent tray. The pump body connector is located above the pump body connection hole, and the snap-fit ​​connector is located above the selector valve. The lifting mechanism can drive the pump body connector to align with the pump body connection hole while the snap-fit ​​connector aligns with the selector valve.

[0010] By adopting the above technical solution, the chip connector is placed on the placement seat, and the reagent tray device is placed in the reagent tray chamber. The pump body connection hole is aligned with the pump body connector above, and the gate valve is aligned with the gate connector above. The pump body connector is driven to connect with the pump body docking hole through the lifting mechanism. At the same time, the driving gate connector is connected to the valve head of the gate valve. The first motor drives the gate connector to rotate and select one of the liquid reservoirs to connect with the chip connector. The injection pump injects the reagent in the liquid reservoir into the reaction chamber inside the chip connector for tissue sample incubation and staining. This automatically realizes the sequential incubation and staining of tissue samples with multiple reagents on the chip connector, thereby improving the preparation efficiency of sequencing samples.

[0011] Preferably, the lifting mechanism includes a second motor and a second lead screw. The second motor is vertically mounted on the support frame, and the second lead screw is coaxially fixed on the output shaft of the second motor. A sliding seat is vertically slidably connected to the support frame via a guide rail. The injection pump and the pump body connector are fixedly mounted on the sliding seat. The second lead screw is threadedly connected to the sliding seat. A connecting seat is vertically slidably mounted on the support frame via a guide rail. The connecting seat is threadedly connected to the second lead screw. The first motor is fixedly mounted on the connecting seat.

[0012] By adopting the above technical solution, the second motor drives the second lead screw to rotate, thereby synchronously moving the sliding seat and the connecting seat up and down, thereby synchronously realizing the up and down movement of the pump body connector and the rotating docking device, thereby synchronously realizing the connection between the selector valve and the clamp connector and the docking between the pump body connector and the pump body connecting hole.

[0013] Preferably, it also includes a tray drive mechanism, which includes a third motor and a horizontal lead screw. The third motor is mounted on the support frame, and the two sides of the reagent tray are horizontally slidably connected to the support frame via guide rails. The output shaft of the third motor is coaxially and fixedly connected to the horizontal lead screw, and the horizontal lead screw is threadedly connected to the reagent tray.

[0014] By adopting the above technical solution, the reagent tray device is placed in the reagent tray compartment. The horizontal lead screw is driven to rotate by the third motor, thereby moving the reagent tray compartment into the support frame, which facilitates the placement of the reagent tray device.

[0015] Preferably, the reagent tray includes a base plate and a seat, the seat being fixedly mounted on one side of the upper surface of the base plate, and the seat is for embedding the reagent tray; a position sensor is provided on the base plate, and a position detection sensor is provided on the support frame; when the position sensor is located at the position detection sensor, the pump body connector is located above the pump body connection hole, and the snap-fit ​​connector is located above the selector valve.

[0016] By adopting the above technical solution, the tray drive mechanism drives the reagent tray to move the reagent tray device. When the position detection sensor detects the position sensing element, the detection sensor controls the third motor to stop working through the controller, so that the pump body connection hole on the reagent tray device is located directly below the pump body connector and the selector valve is located directly below the clamp connector. At the same time, the second motor is controlled to work, thereby improving the precise docking between the pump body connector and the main body connection hole, as well as between the selector valve and the clamp connector.

[0017] Preferably, a barcode scanner is installed facing downwards on the support frame. The barcode scanner is used to scan the identification code of the reagent tray. The barcode scanner is electrically connected to the communication device through a controller.

[0018] By adopting the above technical solution, the reagent tray is scanned and identified by a barcode scanner, thereby automatically entering the information of the reagent tray into the system.

[0019] Preferably, the gate valve is rotatably connected to the reagent tray, the gate valve is provided with a gate channel, the reagent tray is provided with multiple liquid outlet channels, the inlet ports of the multiple liquid outlet channels are respectively located in the corresponding liquid storage tanks, the reagent tray is provided with a connecting channel extending to the mounting base, the liquid outlet port of the gate channel can be connected to the chip connector through the connecting channel, and the liquid inlet port of the gate channel can be connected to the liquid outlet port of one of the liquid outlet channels by rotating the gate valve.

[0020] By adopting the above technical solution, the multiple reservoirs on the reagent tray can hold various reagents. The chip connector is mounted on the mounting base, connecting it to the communication channel. By rotating the selector valve, the desired reservoir is selected and connected to the selector channel via the outlet channel, thus connecting the reservoir to the chip connector. This allows for precise communication of multiple reagents with the chip connector, facilitating accurate management of various reagents for incubating and staining tissue samples within the chip connector. Furthermore, integrating the chip connector and reagent tray into a single unit reduces the number of valves, sensors, and tubing, saving costs.

[0021] Preferably, it also includes a sealing cap and an opening and closing assembly. The sealing cap covers and seals the liquid storage tank. The sealing cap is provided with a plurality of communication holes that communicate with the outside world. Each communication hole is connected to one of the liquid storage tanks.

[0022] The opening and closing assembly is used to open or close the communication hole. The opening and closing assembly includes a mounting cover and a plurality of sealing members. The plurality of sealing members are disposed on the mounting cover. The mounting cover is located above the sealing cover. The sealing members are inserted into the communication hole and can close the communication hole or open the communication hole as the mounting cover moves up or down.

[0023] By adopting the above technical solution, the sealing cap seals multiple liquid storage tanks, thereby preventing external impurities from contaminating the reagents in the liquid storage tanks. When it is necessary to introduce the reagents from the liquid storage tanks into the chip connector, the mounting cap is pressed down to open the connecting hole, thus ensuring that the air pressure in the liquid storage tanks is at a normal state. When it is not necessary to introduce the reagents from the liquid storage tanks into the chip connector, the mounting cap is moved up to block the connecting hole, thereby sealing the liquid storage tanks and preventing the reagents from flowing into the chip connectors, thus precisely controlling the entry of reagents into the chip connectors.

[0024] Preferably, the opening and closing assembly further includes a return spring; the selector valve includes a valve body, a valve head, and a valve core; the valve core is fixedly connected to the valve head; the valve head is rotatably connected to the valve body; the valve body is provided with a through mounting hole; the mounting cover has a clearance opening corresponding to the mounting hole; one end of the valve head extends through the mounting hole to the clearance opening; the selector channel is provided on the valve core; one end of the return spring abuts against the valve body, and the other end abuts against the mounting cover; the valve head is provided with a snap-fit ​​groove for inserting a snap-fit ​​connector; a downward pressing post is vertically arranged on the housing of the first motor, and the downward pressing post is located above the mounting cover.

[0025] By adopting the above technical solution, the lifting device drives the first motor to move the snap-fit ​​connector downwards and insert it into the snap-fit ​​groove of the valve head. Simultaneously, the downward pressure column on the first motor presses down the mounting cover. The first motor drives the snap-fit ​​connector to rotate the valve head, thereby connecting the valve core's selection channel with the desired liquid outlet channel and opening the connecting hole of the sealing component. This simultaneously maintains the air pressure in the storage tank to a state suitable for liquid outlet. This conveniently and accurately connects the storage tank and the chip connector, preventing accidental entry of reagents from the storage tank into the chip connector.

[0026] Preferably, the chip connector includes a top cover, a glass slide, a base, and a sealing ring. The upper surface of the base is provided with two reagent holes spaced apart, and the lower surface of the base is provided with two mating holes, each of which is connected to a corresponding reagent hole.

[0027] The upper surface of the base is provided with a groove for the sealing ring to be embedded. The glass slide is placed on the surface of the sealing ring. The glass slide, the sealing ring, and the base together form the reaction chamber. The two reagent holes are located at the two ends of the reaction chamber. The upper cover is placed on the glass slide and is snapped onto the base by an elastic snap-fit.

[0028] By adopting the above technical solution, a glass slide with tissue sample is placed face down in a sealing ring to form a closed reaction chamber, and the tissue sample is placed in the reaction chamber. Then, the upper cover is placed on the glass slide, and the base is fixed to the upper cover by an elastic snap-fit ​​component, thereby facilitating and quickly assembling the chip connector.

[0029] Preferably, the upper cover is provided with a transparent imaging window at the position corresponding to the reaction chamber.

[0030] By adopting the above technical solution, after the tissue sample is incubated and stained with reagents in the reaction chamber to prepare a sequencing sample, the sequencing sample is scanned and imaged through the imaging window.

[0031] Preferably, the mounting base is provided with a placement slot for placing the chip connector, and the placement slot is provided with two channel connectors; when the chip connector is placed in the placement slot, the two channel connectors are respectively connected to the corresponding mating holes; one of the channel connectors is connected to the connecting channel, and the other channel connector is connected to the pump body connecting hole.

[0032] By adopting the above technical solution, when the chip connector is placed on the placement slot, the two mating holes on the chip connector are connected to the two channel connectors in the placement slot. After the external injection pump is connected to the pump body connection hole, the injection pump, the chip connector, and the liquid storage tank form a passage, so that the reagent in the liquid storage tank is accurately injected into the reaction chamber of the chip connector by the injection pump.

[0033] Preferably, the mounting base is provided with two latches, which are arranged opposite each other on both sides of the placement slot to engage the chip connector located within the placement slot. Each latch includes a latching part and a connecting part. The latching part is integrally connected to the upper end of the connecting part. Unlocking buttons are slidably provided on both opposite sidewalls of the mounting base. The unlocking buttons are rotatably connected to the lower end of the connecting part. The upper side of the connecting part is rotatably connected to the mounting base via a convex shaft. The mounting base also includes a tension spring, one end of which is fixed to the mounting base, and the other end of which is hung on the lower side of the connecting part.

[0034] By adopting the above technical solution, when the chip connector is placed in the placement slot, the two latches on both sides of the placement slot are fixed in the placement slot by the tension of the spring, so that the reagent hole of the chip connector is stably connected with the channel connector. When it is necessary to remove the chip connector from the placement slot, press the unlock button on both sides of the mounting base, so that the two opposite latches rotate outward, and the chip connector can be removed from the placement slot.

[0035] Preferably, the bottom of the placement groove is provided with a movable plate, and the bottom of the movable plate is provided with a spring. The lower end of the spring abuts against the mounting base, and the upper end of the spring abuts against the bottom of the movable plate. Two clearance holes are provided at intervals on the movable plate, and the clearance holes allow the channel connector to extend into the placement groove.

[0036] By adopting the above technical solution, when the unlock button is installed, the movable plate will be pushed upward by the spring force, thereby pushing the chip connector out of the placement slot, making it convenient to remove the chip connector from the placement slot.

[0037] Preferably, the movable plate is provided with a temperature-controlled heating element.

[0038] By adopting the above technical solution, since reagents and tissue samples need to be incubated at different temperatures, heat is transferred to the reaction chamber of the chip connector through a temperature-controlled heating element, thereby accelerating the incubation speed of the tissue samples.

[0039] In summary, this application includes the following beneficial technical effects: the chip connector is placed on the placement seat, the reagent tray device is placed in the reagent tray chamber, the pump body connection hole corresponds to the pump body connector above, and the gate valve corresponds to the gate connector above. The pump body connector is driven to connect with the pump body docking hole through the lifting mechanism, and at the same time, the driving gate connector is connected to the valve head of the gate valve. The first motor drives the gate connector to rotate and select one of the liquid reservoirs to connect with the chip connector. The injection pump injects the reagent in the liquid reservoir into the reaction chamber inside the chip connector for tissue sample incubation and staining, thereby automatically realizing the sequential incubation and staining of tissue samples on the chip connector by multiple reagents, thereby improving the preparation efficiency of sequencing samples. Attached Figure Description

[0040] Figure 1 This is a schematic diagram of the overall structure of the automated tissue sequencing sample processing device in the embodiments of this application.

[0041] Figure 2 This is a rear view of the automated tissue sequencing sample processing device in the embodiments of this application.

[0042] Figure 3 This is a schematic diagram of the structure of the automated tissue sequencing sample processing device without the reagent tray device in the embodiments of this application.

[0043] Figure 4 This is a schematic diagram of the reagent tray device in the embodiments of this application.

[0044] Figure 5 This is an exploded structural diagram of the reagent tray device in an embodiment of this application.

[0045] Figure 6 This is a schematic diagram of the structure of the automated tissue sequencing sample processing device in this application embodiment without the reagent tray device installed, from another perspective.

[0046] Figure 7 This is a schematic diagram of the channel structure inside the reagent tray in an embodiment of this application.

[0047] Figure 8 This is a schematic diagram of the installation structure of the selector valve and the sealing cover in an embodiment of this application.

[0048] Figure 9 This is an exploded structural diagram of the selector valve, mounting cover, and sealing cover in the embodiments of this application.

[0049] Figure 10 This is an exploded view of the chip connector in an embodiment of this application.

[0050] Figure 11 This is an exploded view of the chip connector in another embodiment of this application.

[0051] Figure 12 This is a schematic diagram of the mounting base and chip connector to be installed in the embodiments of this application.

[0052] Figure 13 This is a schematic diagram of the internal structure of the mounting base in an embodiment of this application.

[0053] Figure 14 This is a schematic diagram of the bottom structure of the mounting base in an embodiment of this application.

[0054] Figure 15 This is an exploded structural diagram of the mounting base in an embodiment of this application.

[0055] Explanation of reference numerals in the attached drawings: 1. Support frame; 11. Reagent tray compartment; 111. Base plate; 1111. Position sensor; 1112. Magnetic female connector; 112. Compartment seat; 12. Injection pump; 13. Pump body connector; 14. Rotary docking mechanism; 141. Snap connector; 142. First motor; 143. Downward pressure column; 15. Lifting mechanism; 151. Second motor; 152. Second lead screw; 153. Sliding seat; 154. Connecting seat; 16. Compartment drive mechanism; 161. Third motor; 162. Horizontal lead screw; 17. Position sensor. 18. Detection sensor; 2. Barcode scanner; 3. Reagent tray; 4. Liquid storage tank; 5. Waste liquid tank; 6. Drain hole; 7. Liquid level sensor; 8. Waste liquid plug; 9. Mounting cavity; 10. Liquid outlet channel; 11. Connecting channel; 22. Sealing cover; 23. Connecting hole; 24. Cover; 25. Elastic locking block; 26. Opening and closing assembly; 27. Mounting cover; 28. Displacement port; 28. Convex ring plate; 28. Limiting notch; 28. Ventilation grille; 28. Sealing component; 28. Sealing 2822, Connecting rod; 28221, Connecting notch; 283, Return spring; 29, Slot; 3, Chip connector; 31, Top cover; 32, Glass slide; 321, Clearance hole; 33, Base; 331, Reagent port; 34, Sealing ring; 332, Docking hole; 333, Insert groove; 334, Snap-fit ​​hole; 35, Elastic snap-fit ​​component; 36, Foot pad; 37, O-ring; 38, Imaging window; 4, Selector valve; 41, Selector groove; 42, Valve body; 421, Mounting hole; 422, Insert groove; 423, Fixing block; 43. Valve head; 431. Valve head; 4311. Snap-fit ​​groove; 432. Mounting part; 4321. Mounting groove; 4322. Limiting groove; 44. Valve core; 441. Limiting block; 5. Mounting seat; 51. Pump body connection hole; 52. Placement groove; 53. Channel connector; 54. Snap-fit; 541. Snap-fit ​​part; 542. Connecting part; 543. Unlock button; 544. Tension spring; 55. Movable plate; 551. Clearance hole; 56. Spring; 57. Column; 571. Guide groove; 58. Temperature control heating element; 59. Electrical connector. Detailed Implementation

[0056] The following is in conjunction with the appendix Figure 1-15 This application will be described in further detail.

[0057] This application discloses an automated tissue sequencing sample processing device based on in situ transcriptomics. (Refer to...) Figure 1 The automated tissue sequencing sample processing device includes a support frame 1, a communication device, a reagent tray device, and a reagent tray compartment 11. The reagent tray compartment 11 is fixedly installed at the bottom of the support frame 1 and is used to hold the reagent tray device.

[0058] Combination Figure 1 and Figure 2 The connecting device includes an injection pump 12, a pump body connector 13, a rotary docking mechanism 14, and a lifting mechanism 15. The injection pump 12 and the lifting mechanism 15 are mounted on the support frame 1. Two pump body connectors 13 are provided, and the pump body connectors 13 are connected to the injection pump 12 through pipelines. The lifting mechanism 15 drives the pump body connectors 13 and the rotary docking mechanism 14 to move up and down synchronously.

[0059] Combination Figure 2 The rotating docking mechanism 14 includes a snap-fit ​​connector 141 and a first motor 142. The output shaft of the first motor 142 is fixedly connected to the snap-fit ​​connector 141. Multiple downward pressing columns 143 are vertically fixed on the housing of the first motor 142.

[0060] Combination Figure 3 The lifting mechanism 15 includes a second motor 151 and a second lead screw 152. The second motor 151 is vertically mounted on the support frame 1, and the second lead screw 152 is coaxially fixed on the output shaft of the second motor 151. A sliding seat 153 is vertically slidably connected to the support frame 1 via a guide rail. The injection pump 12 and the pump body connector 13 are fixedly mounted on the sliding seat 153. The second lead screw 152 is threadedly connected to the sliding seat 153. A connecting seat 154 is vertically slidably mounted on the support frame 1 via a guide rail. The connecting seat 154 is threadedly connected to the second lead screw 152. The first motor 142 is fixedly mounted on the connecting seat 154.

[0061] Combination Figure 4 and Figure 5The reagent tray device includes a reagent tray 2, a mounting base 5, a chip connector 3, and a gate valve 4. The mounting base 5 is fixedly mounted on one side of the reagent tray 2. The reagent tray 2 has multiple liquid reservoirs 21 and a waste liquid reservoir 22 spaced apart along its circumference; the reagent tray 2 is circular. The multiple liquid reservoirs 21 store different reagents and buffer solutions, and their volumes can vary as needed. The chip connector 3 has a reaction chamber for incubating tissue samples and is detachably mounted on the mounting base 5. The gate valve 4 is mounted on the reagent tray 2 and can rotate to connect one of the liquid reservoirs 21 to the chip connector 3. The mounting base 5 has a pump connection hole 51, which connects to the chip connector 3.

[0062] Combination Figure 3 and Figure 6 The reagent tray compartment 11 includes a base plate 111 and a seat 112. The seat 112 is fixedly mounted on one side of the upper surface of the base plate 111, and the reagent tray 2 is embedded in the seat 112. It also includes a tray drive mechanism 16, which includes a third motor 161 and a horizontal lead screw 162. The third motor 161 is mounted on a support frame 1. The reagent tray compartment 11 is horizontally slidably connected to the support frame 1 on both sides via guide rails. The output shaft of the third motor 161 is coaxially and fixedly connected to the horizontal lead screw 162, and the horizontal lead screw 162 is threadedly connected to the reagent tray compartment 11. When the reagent tray device is placed inside the reagent tray compartment 11, the third motor 161 drives the horizontal lead screw 162 to rotate, thereby moving the reagent tray compartment 11 into the support frame 1, thus facilitating the placement of the reagent tray device. A position sensor 1111 is installed on the base plate 111, and a position detection sensor 17 is installed on the support frame 1. When the position sensor 1111 is located at the position detection sensor 17, the pump body connector 13 is located above the pump body connection hole 51, and the snap-fit ​​connector 141 is located above the selector valve 4. The tray drive mechanism 16 drives the reagent tray 11 to move the reagent tray device. When the position detection sensor 17 detects the position sensor 1111, the detection sensor controls the third motor 161 to stop working through the controller, so that the pump body connection hole 51 on the reagent tray device is located directly below the pump body connector 13, and the selector valve 4 is located directly below the snap-fit ​​connector 141. At the same time, the second motor 151 is controlled to work, thereby improving the precise docking between the pump body connector 13 and the main body connection hole, and between the selector valve 4 and the snap-fit ​​connector 141.

[0063] Combination Figure 4 and Figure 6 An electrical connector 59 is provided on one side of the mounting base 5, and a magnetic female connector 1112 connected to the electrical connector 59 is provided on the base plate 111.

[0064] Combination Figure 6A barcode scanner 18 is installed facing downwards on the support frame 1. The barcode scanner 18 is used to scan the identification code of the reagent tray 2. The barcode scanner 18 is electrically connected to the communication device through the controller. By scanning the barcode of the reagent tray 2 with the barcode scanner 18, the information of the reagent tray 2 is automatically entered into the system.

[0065] Combination Figure 7 A mounting cavity 23 for accommodating the selector valve 4 is provided in the middle of the reagent tray 2. Multiple liquid outlet channels 24 are provided in the bottom of the reagent tray 2, and the inlet ports of the multiple liquid outlet channels 24 are connected to the liquid storage tank 21. A connecting channel 25 extending to the mounting base 5 is provided in the bottom of the reagent tray 2, and the inlet port of the connecting channel 25 is located in the middle of the bottom of the reagent tray 2. The outlet ports of the multiple liquid outlet channels 24 are distributed at intervals from the inlet port of the connecting channel 25 at a circular distance from each other on the bottom of the reagent tray 2.

[0066] Combination Figure 7 and Figure 8 The bottom of the selector valve 4 is provided with a selector groove 41, forming a selector channel with the bottom of the reagent tray 2. The length of the selector channel is equal to the distance between the inlet port of the connecting channel 25 and the outlet port of the outlet channel 24. The outlet port of the selector channel is connected to the inlet port of the connecting channel 25. The chip connector 3 can be connected to one of the outlet ports of the outlet channel 24 by rotating the selector valve 4. The chip connector 3 is installed on the mounting base 5, so that the chip connector 3 is connected to the connecting channel 25. By rotating the selector valve 4, the required reservoir 21 is selected to be connected to the selector channel through the outlet channel 24, thereby connecting the reservoir 21 to the chip connector 3. This allows for precise connection of multiple reagents to the chip connector 3, facilitating accurate management of multiple reagents for incubating and staining tissue samples in the chip connector 3. Furthermore, integrating the chip connector 3 and the reagent tray 2 into one unit reduces the number of valves, sensors, and tubing used, saving costs.

[0067] Combination Figure 5 and Figure 9It also includes a sealing cap 26, a cap body 27, and an opening and closing assembly 28. The sealing cap 26 covers and seals the liquid storage tank 21 and the waste liquid tank 22, thereby preventing external impurities from contaminating the reagents in the liquid storage tank 21. The sealing cap 26 is provided with multiple connecting holes 261 that communicate with the outside world, and each connecting hole 261 corresponds to a liquid storage tank 21. The opening and closing assembly 28 is used to open or close the connecting hole 261 of the corresponding liquid storage tank 21. The opening and closing assembly 28 includes a mounting cap 281 and multiple sealing elements 282. The multiple sealing elements 282 are disposed on the mounting cap 281, which is located above the sealing cap 26. The sealing elements 282 are inserted into the connecting holes 261 and can close the connecting holes 261 or open the connecting holes 261 as the mounting cap 281 moves upward or downward. The cover 27 has a hollowed-out center to allow room for the installation of the cover 281. The side wall of the cover 27 is provided with an elastic locking block 271, and the side wall of the reagent tray 2 is provided with a locking groove 29. The cover 27 is placed on the reagent tray 2, and the elastic locking block 271 engages with the locking groove 29, thereby fixing the sealing cover 26 on the reagent tray 2.

[0068] Combination Figure 8 and Figure 9 The opening and closing assembly 28 also includes a return spring 283. The selector valve 4 includes a valve body 42, a valve head 43, and a valve core 44, which is fixedly connected to the valve head 43. Specifically, the valve core 44 is disc-shaped, and two limiting blocks 441 are spaced horizontally on the side wall of the valve core 44. The selector groove 41 is located at the bottom of the valve core 44. The valve head 43 includes a mounting part 432 and a valve head 43 part. The bottom of the mounting part 432 is provided with a mounting groove 4321 for mounting the valve core 44, and the mounting part 432 is provided with a limiting groove 4322 for the limiting blocks 441 to be inserted, so that the valve core 44 can rotate with the valve head 43. The diameter of the mounting part 432 is larger than the diameter of the valve head 43 part. The upper part of the valve head 43 part is provided with a snap-fit ​​groove 4311, which has a quincunx cross-sectional area and is inserted into the snap-fit ​​connector 141. The valve body 42 has a through mounting hole 421. The valve head 43 is rotatably connected to the valve body 42. The mounting cover 281 and the sealing cover have clearance openings 2811 at the corresponding positions of the mounting hole 421. One end of the valve head 43 extends through the mounting hole 421 to the clearance opening 2811. The valve body 42 is fixedly mounted on the bottom of the reagent tray 2. One end of the return spring 283 abuts against the valve body 42, and the other end abuts against the mounting cover 281. Specifically, the valve body 42 has a downwardly opening recess 422, and the return spring 283 is embedded in the recess 422. The lower surface of the mounting cover 281 has a downwardly opening convex ring plate 2812 with a limit notch 2813. The outer wall of the valve body 42 has a fixing block 423, which is embedded in the limit notch 2813, thereby restricting the circumferential movement of the mounting cover 281. The mounting cover 281 is provided with a ventilation grille 2814 at the edge of the relief opening 2811.

[0069] The lifting device drives the first motor 142 to move the snap-fit ​​connector 141 downwards and insert it into the snap-fit ​​groove 4311. At the same time, the pressing column 143 on the first motor 142 presses down the mounting cover 281. The first motor 142 drives the snap-fit ​​connector 141 to rotate the valve head 43, thereby driving the valve core 44 to connect with the required liquid outlet channel 24, and simultaneously opening the connecting hole 261 of the sealing component 282. This simultaneously maintains the air pressure in the liquid storage tank 21 to a state that allows liquid to be discharged. This conveniently and accurately connects the liquid storage tank 21 with the chip connector 3, preventing reagents in the liquid storage tank 21 from entering the chip connector due to misoperation.

[0070] Reference Figure 8 and Figure 9 The sealing component 282 includes a sealing buckle 2821 and a connecting rod 2822. One end of the connecting rod 2822 is fixed to the bottom of the mounting cover 281, and the sealing buckle 2821 is detachably fixed to the other end of the connecting rod 2822. A vertical connecting notch 28221 is provided on the side wall of the connecting rod 2822, and multiple connecting notches 28221 are provided along the circumference of the connecting rod 2822. The connecting rod 2822 is movably inserted into the connecting hole 261. The sealing buckle 2821 is located below the mounting cover 281, and the cross-sectional area of ​​the sealing buckle 2821 is larger than the area of ​​the connecting hole 261. The thickness of the mounting cover 281 is smaller than the height of the connecting notch 28221.

[0071] Under the elastic force of the return spring 283, the mounting cover 281 causes the sealing buckle 2821 to abut against the lower surface of the sealing cover 26, thereby closing the connecting hole 261. When the mounting cover 281 is pressed down, since the thickness of the mounting cover 281 is less than the height of the connecting notch 28221 on the side wall of the connecting rod 2822, the connecting hole 261 is opened through the connecting notch 28221, thus facilitating the opening or closing of the connecting hole 261.

[0072] Combination Figure 10 and Figure 11The chip connector 3 includes a top cover 31, a glass slide 32, a base 33, and a sealing ring 34. The upper surface of the base 33 has two reagent holes 331 spaced apart, and the lower surface of the base 33 has two mating holes 332, which communicate with their corresponding reagent holes 331. The upper surface of the base 33 has a groove 333 for the sealing ring 34 to be inserted. The glass slide 32 is placed on the surface of the sealing ring 34. The glass slide 32, the sealing ring 34, and the base 33 together form a reaction chamber. The two reagent holes 331 are located at opposite ends of the reaction chamber. The top cover 31 is placed on the glass slide 32 and is interlocked with the base 33 by an elastic snap-fit ​​member 35. A glass slide 32 containing a tissue sample is placed face down inside the sealing ring 34 to form a closed reaction chamber, with the tissue sample positioned within the chamber. Then, the top cover 31 is placed on the glass slide 32, and the base 33 is then snapped into place by the elastic snap-fit ​​member 35, thus facilitating rapid assembly of the chip connector 3. The base 33 has four rectangular feet 36 embedded at its bottom. An O-ring 37 is embedded in the mating hole 332.

[0073] Multiple elastic locking elements 35 are provided, spaced apart on the lower surface of the upper cover 31. The base 33 has multiple locking holes 334, and the glass slide 32 has clearance holes 321. The elastic locking elements 35 pass through the clearance holes 321 and lock onto the locking holes 334. By having multiple elastic locking elements 35 on the upper cover 31 pass through the clearance holes 321 and lock onto the locking holes 334 on the base 33, the stability of the glass slide 32 placed between the base 33 and the upper cover 31 is improved, thereby forming a stable reaction chamber. In this embodiment, four elastic locking elements 35 are provided, and the four elastic locking elements 35 are arranged in a rectangular shape.

[0074] A transparent imaging window 38 is provided on the top cover 31 corresponding to the position of the reaction chamber. After the tissue sample is incubated and stained with reagents in the reaction chamber to prepare a sequencing sample, the sequencing sample is scanned and imaged through the imaging window 38.

[0075] Combination Figure 7 and Figure 12The mounting base 5 is provided with a placement slot 52 for placing the chip connector 3. Two channel connectors 53 are provided within the placement slot 52. When the chip connector 3 is placed in the placement slot 52, the two channel connectors 53 are respectively connected to their corresponding mating holes 332. In this embodiment, two pump body connectors 13 are provided; one channel connector 53 is connected to the connecting channel 25, and the other channel connector 53 is connected to one of its pump body connection holes 51. While the chip connector 3 is placed in the placement slot 52, the two mating holes 332 on the chip connector 3 are connected to the two channel connectors 53 in the placement slot 52. After the external injection pump 12 is connected to the pump body connection hole 51, a passage is formed between the injection pump 12, the chip connector 3, and the storage tank 21, thereby allowing the injection pump 12 to precisely inject the reagent in the storage tank 21 into the reaction chamber of the chip connector 3. The waste liquid tank 22 is connected to another pump body connection hole 51. A drain hole 221 is provided at the bottom of the reagent tray 2, located at the bottom of the waste liquid tank 22. After one of the reagents has incubated and stained the tissue sample in the reaction chamber, the waste liquid after washing the tissue sample in the reaction chamber with the buffer solution in the storage tank 21 is discharged into the waste liquid tank 22 through the syringe pump 12.

[0076] Combination Figure 12 and Figure 13 The mounting base 5 is equipped with two latches 54, which are positioned opposite each other on both sides of the placement slot 52 to engage the chip connector 3 located within the placement slot 52. Each latch 54 includes a latching part 541 and a connecting part 542. The latching part 541 is integrally connected to the upper end of the connecting part 542. Unlocking buttons 543 are slidably mounted on both opposite sidewalls of the mounting base 5. The unlocking buttons 543 are rotatably connected to the lower end of the connecting part 542, and the upper side of the connecting part 542 is rotatably connected to the mounting base 5 via a convex shaft. A liquid level sensor 222 for detecting waste liquid in the waste liquid tank 22 is installed on the sidewall of the reagent tray 2. When the liquid level sensor 222 detects excessive waste liquid in the waste liquid tank 22, it promptly cleans the waste liquid.

[0077] Combination Figure 13 and Figure 14It also includes a tension spring 544, one end of which is fixed to the mounting base 5, and the other end is hung on the lower side of the connecting part 542. Two tension springs 544 are provided, located on both sides of the unlocking button 543. When the chip connector 3 is placed in the placement slot 52, the two latches 54 on both sides of the placement slot 52 are fixed in place by the snap-fit ​​parts 541 of the latches 54 under the tension of the tension springs 544, thereby ensuring that the reagent hole 331 of the chip connector 3 is stably connected to the channel connector 53. When it is necessary to remove the chip connector 3 from the placement slot 52, the unlocking button 543 on both sides of the mounting base 5 is pressed, causing the two opposing snap-fit ​​parts 541 to rotate outward, thereby removing the chip connector 3 from the placement slot 52. A waste liquid plug 223 is inserted into the drain hole 221. By opening the waste liquid plug 223, the waste liquid in the waste liquid pool 22 is discharged from the drain hole 221 in a timely manner.

[0078] Combination Figure 15 A movable plate 55 is provided at the bottom of the placement groove 52, and a spring 56 is provided at the bottom of the movable plate 55. The lower end of the spring 56 abuts against the mounting base 5, and the upper end of the spring 56 abuts against the bottom of the movable plate 55. Two clearance holes 551 are provided on the movable plate 55 at intervals, and the clearance holes 551 allow the channel connector 53 to extend into the placement groove 52. A column 57 is provided in the mounting base 5, and the column 57 is provided with a guide groove 571, in which the spring 56 is placed.

[0079] When the unlock button 543 is installed, the movable plate 55 is pushed upward by the spring force of the spring 56, thereby pushing the chip connector 3 out of the placement slot 52, so that the chip connector 3 can be easily removed from the placement slot 52.

[0080] A temperature-controlled heating element 58 is provided on the movable plate 55. In this embodiment, the temperature-controlled heating element 58 is a semiconductor cooler. Since the reagents and tissue samples need to be incubated at different temperatures, the temperature-controlled heating element 58 transfers heat to the reaction chamber of the chip connector 3, thereby accelerating the incubation speed of the tissue samples.

[0081] The implementation principle of an automated tissue sequencing sample processing device based on in situ transcriptomics in this application embodiment is as follows: the chip connector 3 is placed on the placement seat, the reagent tray device is placed in the reagent tray chamber 11, the pump body connection hole 51 is aligned with the pump body connector 13 above, and the gate valve 4 is aligned with the gate connector 141 above. The lifting mechanism 15 drives the pump body connector 13 to connect with the pump body docking hole 332, and at the same time connects the gate connector 141 to the valve head 43 of the gate valve 4. The first motor 142 drives the gate connector 141 to rotate the gate valve 4 to select one of the liquid reservoirs 21 to connect with the chip connector 3. The injection pump 12 injects the reagent in the liquid reservoir 21 into the reaction chamber in the chip connector 3 for tissue sample incubation and staining, thereby automatically realizing the sequential incubation and staining of tissue samples in the chip connector 3 by multiple reagents, thereby improving the preparation efficiency of sequencing samples.

[0082] The above are all preferred embodiments of this application, and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, shape and principle of this application should be covered within the scope of protection of this application.

Claims

1. An automated tissue sequencing sample processing device based on in situ transcriptomics, characterized in that, include: Support frame (1); A connecting device is provided, comprising an injection pump (12), a pump body connector (13), a rotary docking mechanism (14), and a lifting mechanism (15). The injection pump (12) and the lifting mechanism (15) are mounted on a support frame (1). The pump body connector (13) is connected to the injection pump (12) via a pipeline. The lifting mechanism (15) drives the pump body connector (13) and the rotary docking mechanism (14) to move up and down synchronously. The rotary docking mechanism (14) includes a snap-fit ​​connector (141) and a first motor (142), and the first motor (142) drives the snap-fit ​​connector (141) to rotate. A reagent tray device includes a reagent tray (2), a mounting base (5), a chip connector (3), and a gate valve (4). The mounting base (5) is fixedly disposed on one side of the reagent tray (2). The reagent tray (2) has multiple liquid reservoirs (21) spaced apart along the circumference. The chip connector (3) has a reaction chamber for incubating tissue samples. The chip connector (3) is detachably mounted on the mounting base (5). The gate valve (4) is disposed on the reagent tray (2) and can rotate to select one of the liquid reservoirs (21) to communicate with the chip connector (3). The mounting base (5) is provided with a pump body connection hole (51), which communicates with the chip connector (3). The reagent tray (11) is located on the lower side of the support frame (1). The reagent tray device is placed on the reagent tray (11). The pump body connector (13) is located above the pump body connection hole (51). The snap connector (141) is located above the selector valve (4). The lifting mechanism (15) can drive the pump body connector (13) to dock with the pump body connection hole (51) while the snap connector (141) docks with the selector valve (4). It also includes a sealing cap (26) and an opening and closing assembly (28). The sealing cap (26) covers and seals the liquid storage tank (21). The sealing cap (26) is provided with a plurality of communication holes (261) that communicate with the outside world. Each communication hole (261) is connected to one of the liquid storage tanks (21). The opening and closing assembly (28) is used to open or close the connecting hole (261). The opening and closing assembly (28) includes a mounting cover (281) and a plurality of sealing members (282). The plurality of sealing members (282) are disposed on the mounting cover (281). The mounting cover (281) is located above the sealing cover (26). The sealing members (282) are inserted into the connecting hole (261) and can close the connecting hole (261) or open the connecting hole (261) as the mounting cover (281) moves upward; The opening and closing assembly (28) further includes a return spring (283). The selector valve (4) includes a valve body (42), a valve head (43), and a valve core (44). The valve core (44) is fixedly connected to the valve head (43), and the valve head (43) is rotatably connected to the valve body (42). The valve body (42) is provided with a through mounting hole. The mounting cover (281) is provided with a clearance opening (2811) corresponding to the mounting hole. One end of the valve head (43) passes through the... The mounting hole extends to the clearance port (2811), and the selection channel is provided on the valve core (44); one end of the return spring (283) abuts against the valve body (42), and the other end abuts against the mounting cover (281); the valve head (43) is provided with a snap-fit ​​groove (4311) for inserting the snap-fit ​​connector (141); a downward pressing column (143) is vertically arranged on the housing of the first motor (142), and the downward pressing column (143) is located above the mounting cover (281).

2. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 1, characterized in that: The lifting mechanism (15) includes a second motor (151) and a second lead screw (152). The second motor (151) is vertically mounted on the support frame (1). The second lead screw (152) is coaxially fixed on the output shaft of the second motor (151). A sliding seat (153) is vertically slidably connected to the support frame (1) via a guide rail. The injection pump (12) and the pump body connector (13) are fixedly mounted on the sliding seat (153). The second lead screw (152) is threadedly connected to the sliding seat (153). A connecting seat (154) is vertically slidably connected to the support frame (1) via a guide rail. The connecting seat (154) is threadedly connected to the second lead screw (152). The first motor (142) is fixedly mounted on the connecting seat (154).

3. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 1, characterized in that: It also includes a tray drive mechanism (16), which includes a third motor (161) and a horizontal lead screw (162). The third motor (161) is mounted on the support frame (1). The reagent tray (11) is horizontally slidably connected to the support frame (1) on both sides via guide rails. The output shaft of the third motor (161) is coaxially fixedly connected to the horizontal lead screw (162). The horizontal lead screw (162) is threadedly connected to the reagent tray (11).

4. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 3, characterized in that: The reagent tray compartment (11) includes a base plate (111) and a seat (112). The seat (112) is fixedly installed on one side of the upper surface of the base plate (111) and the reagent tray (2) is embedded in the seat (112). A position sensing element (1111) is provided on the base plate (111), and a position detection sensor (17) is provided on the support frame (1). When the position detection sensor (17) detects the position sensing element (1111), the pump body connector (13) is located above the pump body connection hole (51), and the snap connector (141) is located above the selector valve (4).

5. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 1, characterized in that: The support frame (1) is provided with a barcode scanner (18) facing downwards. The barcode scanner (18) is used to scan the identification code of the reagent tray (2). The barcode scanner (18) is electrically connected to the communication device through the controller.

6. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 1, characterized in that: The selector valve (4) is rotatably connected to the reagent tray (2). The selector valve (4) is provided with a selector channel. The reagent tray (2) is provided with multiple liquid outlet channels (24). The inlet ports of the multiple liquid outlet channels (24) are respectively located in the corresponding liquid storage tank (21). The reagent tray (2) is provided with a connecting channel (25) extending to the mounting base (5). The liquid outlet port of the selector channel can be connected to the chip connector (3) through the connecting channel (25). The liquid inlet port of the selector channel can be connected to the liquid outlet port of one of the liquid outlet channels (24) by rotating the selector valve (4).

7. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 1, characterized in that: The chip connector (3) includes a top cover (31), a glass slide (32), a base (33) and a sealing ring (34). The upper surface of the base (33) is provided with two reagent holes (331) spaced apart, and the lower surface of the base (33) is provided with two docking holes (332). The two docking holes (332) are respectively connected to the corresponding reagent holes (331). The upper surface of the base (33) is provided with a groove (333) for the sealing ring (34) to be inserted. The glass slide (32) is placed on the surface of the sealing ring (34). The glass slide (32), the sealing ring (34) and the base (33) together form the reaction chamber. The two reagent holes (331) are located at the two ends of the reaction chamber respectively. The upper cover (31) is placed on the glass slide (32). The upper cover (31) is interlocked with the base (33) by an elastic snap-fit ​​(35).

8. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 7, characterized in that: The upper cover (31) is provided with a transparent imaging window (38) corresponding to the position of the reaction chamber.

9. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 7, characterized in that: The mounting base (5) is provided with a placement slot (52) for placing the chip connector (3). Two channel connectors (53) are provided in the placement slot (52). When the chip connector (3) is placed in the placement slot (52), the two channel connectors (53) are respectively connected to the corresponding docking holes (332). One of the channel connectors (53) is connected to the connecting channel (25), and the other channel connector (53) is connected to the pump body connecting hole (51).

10. An automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 9, characterized in that: The mounting base (5) is provided with buckles (54), and there are two buckles (54). The two buckles (54) are arranged opposite each other on both sides of the placement groove (52) for snapping the chip connector (3) into the placement groove (52). The buckle (54) includes a snap-fit ​​part (541) and a connecting part (542). The snap-fit ​​part (541) is integrally connected to the upper end of the connecting part (542). The two opposite side walls of the mounting base (5) are slidably provided with unlocking buttons (543). The unlocking buttons (543) are rotatably connected to the lower end of the connecting part (542). The upper side of the connecting part (542) is rotatably connected to the mounting base (5) through a convex shaft. It also includes a tension spring (544). One end of the tension spring (544) is fixed on the mounting base (5), and the other end of the tension spring (544) is hung on the lower side of the connecting part (542).

11. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 10, characterized in that: The bottom of the placement slot (52) is provided with a movable plate (55), and the bottom of the movable plate (55) is provided with a spring (56). The lower end of the spring (56) abuts against the mounting base (5), and the upper end of the spring (56) abuts against the bottom of the movable plate (55). Two clearance holes (551) are provided on the movable plate (55) at intervals. The clearance holes (551) allow the channel connector (53) to extend into the placement slot (52).

12. The automated tissue sequencing sample processing device based on in situ transcriptomics according to claim 11, characterized in that: A temperature-controlled heating element (58) is provided on the movable plate (55).

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