A swing device for an in situ RNA sequencer

Abstract

This application relates to the field of fluorescence in situ sequencing technology equipment, and in particular to a swing device for an in situ RNA sequencer. The device includes a swing platform rotatably mounted on a base, the swing platform for holding a chip adapter, and a sample inlet / outlet connector. When the chip adapter is placed on the swing platform, the sample inlet / outlet connector communicates with the chip adapter. A driving member, connected to the swing platform, drives the swing platform to tilt and intermittently reciprocate in the direction of sample inlet or outlet. This application has the advantage of improving the uniformity and efficiency of tissue sample incubation and staining.

Description

Technical Field

[0001] This application relates to the field of fluorescence in situ sequencing technology equipment, and in particular to a swing device for an in situ RNA sequencer. 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] Based on the aforementioned fluorescence in situ sequencing technology, staining reagents are injected into a chip adapter containing tissue samples via a syringe pump for incubation and staining. However, existing methods of injecting reagents into the chip adapter via a syringe pump suffer from uneven tissue sample incubation and low efficiency. Summary of the Invention

[0005] To improve the uniformity and efficiency of tissue sample incubation and staining, this application provides a swing device for an in situ RNA sequencer.

[0006] The swing device for an in situ RNA sequencer provided in this application adopts the following technical solution: A swing device for an in situ RNA sequencer, comprising: A rotatable platform is rotatably mounted on a base body. The platform is used to place a chip adapter and is equipped with a sample inlet / outlet connector. When the chip adapter is placed on the platform, the sample inlet / outlet connector can communicate with the chip adapter. A driving component, which is connected to a swing platform, is used to drive the swing platform to tilt and reciprocate intermittently in the direction of sample feeding or discharging from the chip adapter.

[0007] By adopting the above technical solution, the chip adapter is placed on the stage, and the chip adapter is connected to the sample inlet and outlet connectors on the stage to inject reagents. During the process of reagents entering the chip adapter, the stage is driven by the driving component to tilt and swing the chip adapter, thereby realizing the dynamic mixing of tissue samples and reagents in the chip adapter, accelerating the full reaction of reagents and tissue samples, and minimizing the incubation time.

[0008] Preferably, the stage base is fixedly provided with a channel side block, a fixed side block, and a limiting side block. The channel side block and the fixed side block are arranged opposite to each other, and the limiting side block is connected between the channel side block and the fixed side block. The channel side block, the fixed side block, and the limiting side block together form a placement groove for placing a chip adapter. The sample inlet / outlet connector is connected to the channel side block, and the side wall of the channel side block is provided with a sample inlet / outlet connection hole. When the chip adapter is located in the placement groove, the sample inlet / outlet connection hole is connected to the chip adapter.

[0009] By adopting the above technical solution, the chip adapter is placed in the placement slot and connected to the chip adapter through the sample inlet / outlet connection hole on the channel side block, thereby realizing the stable entry and exit of reagents while the chip adapter swings.

[0010] Preferably, a clamping assembly is provided on the fixed side block. The clamping assembly includes a clamping block, a spring, and a cam handle. The fixed side block has a through hole through which the clamping block moves. One end of the screw portion of the cam handle is fixedly connected to the fixed side block. The spring is sleeved on the screw portion of the cam handle. One end of the spring abuts against the fixed side block, and the other end abuts against the clamping block. The handle portion of the cam handle abuts against the clamping block.

[0011] By adopting the above technical solution, the chip adapter is placed in the placement slot. By rotating the cam handle, the clamping block abuts against the side wall of the chip adapter, thereby improving the stability of the chip adapter in the placement slot and ensuring stable connection between the chip adapter and the sample inlet / outlet connection hole.

[0012] Preferably, the sidewall of the limiting side block is provided with a magnetic element for engaging with the chip adapter.

[0013] By adopting the above technical solution, the chip adapter is placed on the placement slot and pre-positioned and fixed by magnetic components, thereby facilitating the rapid docking of the chip adapter with the sample inlet / outlet docking hole.

[0014] Preferably, the surface of the swing platform is provided with a heating plate.

[0015] By adopting the above technical solution, the heating plate on the stage conducts heat to the chip adapter, thereby accelerating the incubation time of the tissue sample.

[0016] Preferably, the swing platform located in the placement slot is provided with a proximity switch for detecting whether a chip adapter is placed.

[0017] By adopting the above technical solution, a proximity switch is used to detect whether a chip adapter is placed, and the detection signal from the proximity switch is used to control the sample output from the sample inlet / outlet connector.

[0018] Preferably, the driving component is a motor, and the output shaft of the motor is connected to the swing platform base.

[0019] By adopting the above technical solution, the pendulum platform can be easily driven to swing back and forth by a motor.

[0020] Preferably, the bottom of the swing platform is provided with a clearance slope from the middle to both sides, and a trigger is provided on the clearance slope. The base is provided with a detection element for detecting the trigger, and the detection element controls the operation of the motor through a controller.

[0021] By adopting the above technical solution, the tilt of the pendulum platform during the swing driven by the motor is controlled by the trigger on the inclined surface and the detection device on the platform.

[0022] In summary, this application includes at least one of the following beneficial technical effects: the chip adapter is placed on the stage, and the chip adapter is connected to the sample inlet / outlet connector on the stage for reagent injection. During the process of reagent entering the chip adapter, the stage is driven by a driving component to tilt and swing the chip adapter, thereby realizing dynamic mixing of tissue sample and reagent in the chip adapter, accelerating the rapid distribution of reagent in the reaction chamber and achieving full contact and reaction with tissue sample, thus minimizing the incubation time. Attached Figure Description

[0023] Figure 1 This is a schematic diagram of the overall structure of the in situ RNA sequencer in the embodiments of this application.

[0024] Figure 2 This is a schematic diagram of the chip adapter to be assembled in the swing device in the embodiments of this application.

[0025] Figure 3 This is a schematic diagram of the structure of the swing device in the embodiments of this application.

[0026] Figure 4 This is an exploded structural diagram of the clamping assembly on the fixed side block in an embodiment of this application.

[0027] Explanation of reference numerals in the attached drawings: 1. Base; 11. Bearing seat; 12. Detector; 2. Reagent mounting tray; 3. Chip adapter; 31. Magnetic component; 4. Slab base; 41. Sample inlet / outlet connector; 42. Channel side block; 421. Sample inlet / outlet connection hole; 422. O-ring; 43. Fixing side block; 431. Through hole; 432. Mounting block; 44. Limiting side block; 441. Magnetic suction component; 45. Placement slot; 46. Clearance slope; 47. Trigger; 48. Proximity switch; 49. Heating plate; 5. Drive component; 6. Clamping assembly; 61. Clamping block; 611. Clearance mounting slot; 62. Spring; 63. Cam handle. Detailed Implementation

[0028] The following is in conjunction with the appendix Figure 1-4 This application will be described in further detail.

[0029] This application discloses a swing device for an in situ RNA sequencer. (See also...) Figure 1 and Figure 2 The in situ RNA sequencer includes a base 1, a reagent mounting tray 2, a chip adapter 3, and a swaying device. Both the reagent mounting tray 2 and the swaying device are mounted on the base 1. The chip adapter 3 contains a reaction chamber for tissue samples. The chip adapter 3 is placed on the swaying device and connected to reagent tubes on the reagent mounting tray 2 via tubing. A syringe pump injects reagents from the reagent tubes into the reaction chamber of the chip adapter 3 for incubation and staining. The swaying device, rotatably mounted on the base 1 of the sequencer, tilts the chip adapter 3 intermittently in the direction of sample entry or exit from the reaction chamber, accelerating reagent entry into the reaction chamber and rapidly removing air bubbles. This achieves dynamic mixing of the tissue sample and reagents, reducing the incubation and staining time.

[0030] Reference Figure 2 and Figure 3 The swing device includes a swing platform 4 and a drive component 5. A bearing seat 11 is provided on the base 1, and the swing platform 4 is rotatably connected to the bearing seat 11 via a bearing. The swing platform 4 is used to place the chip adapter 3 and is provided with a sample inlet / outlet connector 41. When the chip adapter 3 is placed on the swing platform 4, the sample inlet / outlet connector 41 can communicate with the reaction chamber of the chip adapter 3. The drive component 5 is connected to the swing platform 4 and is used to drive the swing platform 4 to tilt and intermittently reciprocate in the direction of sample inlet or outlet of the reaction chamber. By driving the swing platform 4 to tilt and swing the chip adapter 3 through the drive component 5, dynamic mixing of tissue samples and reagents within the chip adapter 3 is achieved, accelerating the rapid distribution of reagents into the reaction chamber to achieve sufficient contact and reaction with the tissue samples, thereby minimizing incubation time. In this embodiment, the drive component 5 is a motor, which is mounted on the base 1. In other embodiments, the drive component 5 can be a combination of a cylinder and a crank-connecting rod assembly to achieve the reciprocating swing of the swing platform 4.

[0031] To improve the stability of the chip adapter 3 placement and facilitate communication with the sample inlet / outlet connectors, the stage 4 is fixedly equipped with a channel side block 42, a fixed side block 43, and a limiting side block 44. The channel side block 42 and the fixed side block 43 are positioned opposite each other on the two side walls of the stage 4. The limiting side block 44 is connected to the side wall of the stage 4 between the channel side block 42 and the fixed side block 43. The channel side block 42, the fixed side block 43, and the limiting side block 44 together form a placement slot 45 for placing the chip adapter 3. Two sample inlet / outlet connectors are provided, with two sample inlet / outlet connectors 41 connected to the channel side block 42. Two sample inlet / outlet connection holes 421 are provided on the side wall of the channel side block 42 facing the fixed side block 43. The sample inlet / outlet connectors communicate with the sample inlet / outlet connection holes 421, with one connector for sample intake and the other for sample output. An O-ring 422 is embedded at the opening of the sample inlet / outlet connection hole 421. With the chip adapter 3 in the placement slot 45, it is connected to the chip adapter 3 through the sample inlet / outlet connection hole 421.

[0032] The bottom of the swing platform 4 is inclined upwards from the middle to both sides, with a clearance slope 46. A trigger 47 is provided on the clearance slope 46, and a detection element 12 for detecting the trigger 47 is provided on the base body 1. The detection element 12 controls the operation of the motor through a controller. During the swinging process driven by the motor, the reciprocating swing angle of the swing platform 4 is controlled by the trigger 47 on the clearance slope 46 and the detection element 12 on the base body 1 through the controller. In this embodiment, the detection element 12 is an infrared sensor.

[0033] The swing base 4 located in the placement slot 45 is equipped with a proximity switch 48 for detecting whether the chip adapter 3 is placed there. By detecting whether the chip adapter 3 is placed by the proximity switch 48, the operation of the drive unit 5 is controlled to avoid accidental operation.

[0034] A magnetic element 441 is embedded in the limiting side block 44, and a magnetic element 31 that attracts the magnetic element 441 is embedded in the side wall of the chip adapter 3. The chip adapter 3 is placed on the placement slot 45 and pre-positioned and fixed by the magnetic element 31, thereby quickly connecting the chip adapter 3 with the sample inlet / outlet docking hole.

[0035] A heating plate 49 is provided on the surface of the stage 4 located in the placement slot 45. The chip adapter 3 placed in the placement slot 45 conducts heat to the reaction chamber through the heating plate 49, thereby accelerating the incubation time of the tissue sample in the reaction chamber.

[0036] Reference Figure 3 and Figure 4To further improve the stability of the chip adapter 3 placed in the placement slot 45 and the stability of its connection with the sample inlet / outlet docking hole, a clamping assembly 6 is provided on the fixed side block 43. The clamping assembly 6 includes a clamping block 61, a spring 62, and a cam handle 63. The fixed side block 43 has a through hole 431, and a mounting block 432 is fixedly installed in the middle of the through hole 431. The clamping block 61 has a clearance mounting groove 611 in the middle. The clamping block 61 moves through the through hole 431, and the mounting block 432 is embedded in the clearance mounting groove 611. One end of the screw part of the cam handle 63 is fixedly connected to the mounting block 432. The spring 62 is sleeved on the screw part of the cam handle 63. One end of the spring 62 abuts against the side wall of the mounting block 432, and the other end abuts against the clearance mounting groove 611 of the clamping block 61. The handle part of the cam handle 63 abuts against the clamping block 61. The chip adapter 3 is placed in the placement slot 45. By rotating the cam handle 63, the clamping block 61 abuts against the side wall of the chip adapter 3, thereby improving the stability of the chip adapter 3 in the placement slot 45 and making the chip adapter 3 stably dock with the sample inlet / outlet connection hole 421.

[0037] The implementation principle of the swing device for an in situ RNA sequencer in this application embodiment is as follows: the chip adapter 3 is placed on the swing base 4, and the chip adapter 3 is connected to the sample inlet / outlet port on the swing base 4 by the clamping component 6. During the process of reagent entering the reaction chamber of the chip adapter 3, the swing base 4 is driven by the driving component 5 to tilt and swing the chip adapter 3. This accelerates the entry of reagent into the reaction chamber of the chip adapter 3, and at the same time quickly removes air bubbles in the reaction chamber, realizing dynamic and uniform mixing of tissue samples and reagents, and reducing the incubation and staining time of tissue samples and reagents.

[0038] The chip adapter 3 is placed on the stage 4 and connected to the sample inlet / outlet connector 41. When the reagent is injected into the reaction chamber by the syringe pump, the drive unit 5 drives the stage 4 to swing and tilt the chip adapter 3 in the sample injection direction, so that the reagent gradually spreads into the entire reaction chamber, thereby reducing the generation of bubbles in the reaction chamber. After the reagent injection is completed, the drive unit 5 drives the stage 4 to swing the chip adapter back and forth, realizing the dynamic mixing of tissue sample and reagent in the chip adapter, accelerating the full reaction of reagent and tissue sample, and minimizing the incubation time. When the reagent is discharged from the reaction chamber by the syringe pump, the drive unit 5 drives the stage 4 to swing and tilt the chip adapter 3 in the sample outlet direction, thereby accelerating the discharge of reagent and reducing the residue of reagent in the reaction chamber.

[0039] 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. A swing device for an in situ RNA sequencer, characterized in that, include: A rotatable stand (4) is rotatably mounted on a base (1). The rotatable stand (4) is used to place a chip adapter (3). The rotatable stand (4) is provided with a sample inlet / outlet connector (41). When the chip adapter (3) is placed on the rotatable stand (4), the sample inlet / outlet connector (41) can communicate with the chip adapter (3). The driving component (5) is connected to the swing base (4) and is used to drive the swing base (4) to drive the chip adapter (3) to tilt and swing intermittently in the direction of sample feeding or sample output of the chip adapter (3). The platform base (4) is fixedly provided with a channel side block (42), a fixed side block (43) and a limiting side block (44). The channel side block (42) and the fixed side block (43) are arranged opposite to each other. The limiting side block (44) is connected between the channel side block (42) and the fixed side block (43). The channel side block (42), the fixed side block (43) and the limiting side block (44) together form a placement groove (45) for placing the chip adapter (3). The sample inlet / outlet connector (41) is connected to the channel side block (42). The side wall of the channel side block (42) is provided with a sample inlet / outlet connection hole (421). When the chip adapter (3) is located in the placement groove (45), the sample inlet / outlet connection hole (421) is connected to the chip adapter (3). A clamping assembly (6) is provided on the fixed side block (43). The clamping assembly (6) includes a clamping block (61), a spring (62), and a cam handle (63). The fixed side block (43) has a through hole (431). The clamping block (61) moves through the through hole (431). One end of the screw part of the cam handle (63) is fixedly connected to the fixed side block (43). The spring (62) is sleeved on the screw part of the cam handle (63). One end of the spring (62) abuts against the fixed side block (43), and the other end abuts against the clamping block (61). The handle part of the cam handle (63) abuts against the clamping block (61).

2. The swing device for an in situ RNA sequencer according to claim 1, characterized in that: The sidewall of the limiting side block (44) is provided with a magnetic element (31) for engaging with the chip adapter (3).

3. The swing device for an in situ RNA sequencer according to claim 1, characterized in that: A heating plate (49) is provided on the surface of the swing stand (4).

4. The swing device for an in situ RNA sequencer according to claim 1, characterized in that: The pedestal (4) located in the placement slot (45) is equipped with a proximity switch (48) for detecting whether the chip adapter (3) is placed.

5. The swing device for an in situ RNA sequencer according to claim 1, characterized in that: The driving component (5) is a motor, and the output shaft of the motor is connected to the swing platform base (4).

6. The swing device for an in situ RNA sequencer according to claim 5, characterized in that: The bottom of the swing platform (4) is inclined upward from the middle to both sides with a clearance slope (46). A trigger (47) is provided on the clearance slope (46). A detection element (12) for detecting the trigger (47) is provided on the seat (1). The detection element (12) controls the operation of the motor through the controller.

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