A chip adapter for an in situ RNA sequencer

Abstract

The application relates to the field of fluorescent in-situ sequencing technology equipment, in particular to a chip adapter of an in-situ RNA sequencer, which comprises a channel seat body internally provided with a first channel and a second channel; the channel seat body is provided with a first docking interface and a second docking interface; the channel seat body is provided with a first sample port and a second sample port; a sealing film is provided with a cavity port, the sealing film is placed on the surface of the channel seat body, the first sample port and the second sample port are distributed on the two end edges in the cavity port; the sealing film, the channel seat body and a slide form a reaction cavity; a cover is used for covering the surface of the sealing film; a buckle assembly is connected with the cover and can be clamped on the channel seat body. The application has the advantages of reducing pollution in the process of capturing and staining the tissue sample and improving the accuracy of detection.

Description

Technical Field

[0001] This application relates to the field of fluorescence in situ sequencing technology equipment, and in particular to a chip adapter 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, sequencing sample preparation requires incubation and staining of tissue slides. However, current methods typically involve manually applying reagents directly onto a glass slide containing the tissue sample, which can easily lead to tissue contamination and affect the accuracy of the sequencing results. Summary of the Invention

[0005] To reduce tissue sample contamination and improve the accuracy of sequencing samples, this application provides a chip adapter for an in situ RNA sequencer.

[0006] The chip adapter for an in situ RNA sequencer provided in this application adopts the following technical solution: A chip adapter for an in situ RNA sequencer, comprising:

[0007] A channel seat body, wherein a first channel and a second channel are provided inside the channel seat body; one surface of the channel seat body is provided with a first pair of interfaces and a second pair of interfaces, one end of the first channel is connected to the first pair of interfaces, and one end of the second channel is connected to the second pair of interfaces; the other surface of the channel seat body is provided with a first sample port and a second sample port, the other end of the first channel is connected to the first sample port, and the other end of the second channel is connected to the second sample port.

[0008] A sealing sheet is provided with a cavity opening. The sealing sheet is placed on the surface of the channel seat. The first sample opening and the second sample opening are distributed at both ends of the cavity opening. The sealing sheet, the channel seat, and the glass slide form a reaction cavity.

[0009] A cover body, the cover body being used to cover a glass slide on the surface of a sealing film;

[0010] A snap-fit ​​assembly, which is connected to the cover and can be snapped onto the channel seat.

[0011] By adopting the above technical solution, a sealing film is first placed on the surface of the channel seat. By adjusting the position of the sealing film, the first sample port and the second sample port are located at opposite ends of the cavity opening. Then, a glass slide is placed on the sealing film to form a reaction cavity. The position of the glass slide is adjusted horizontally so that the tissue sample on the glass slide is located inside the cavity opening. Then, the cover is placed on the glass slide and fixed to the channel seat by a snap-fit ​​assembly. The cover is connected to a multi-port selection valve through the first pair of interfaces and to a syringe pump through the second pair of interfaces. Thus, the reagent in the reagent tube is injected into the reaction cavity for tissue sample incubation and staining. Therefore, contamination during the tissue sample incubation and staining process is reduced, and the accuracy of sequencing sample preparation is improved.

[0012] Preferably, the cover has a transparent window at the position corresponding to the cavity opening, and the thickness of the transparent window is less than the thickness of the cover.

[0013] By adopting the above technical solution, the transparent window allows for convenient observation of the reagents within the reaction chamber, and also enables direct scanning and imaging of the tissue sample after the reaction. In addition, to accelerate the reaction speed between the tissue sample and the reagents, heating is required, and the transparent window allows for rapid heat transfer into the reaction chamber.

[0014] Preferably, the surface of the cover is inclined towards the direction of the transparent window.

[0015] By adopting the above technical solution, the periphery of the cover surface is tilted towards the direction of the transparent window, thereby forming a larger observation angle, which facilitates the observation of the reagent injection in the reaction chamber and facilitates scanning imaging.

[0016] Preferably, the surface of the cover facing the channel seat is provided with a limiting groove, the surface of the channel seat is provided with a mating boss, the sealing film is placed on the mating boss, the first sample port and the second sample port are distributed at intervals along the length direction on the mating boss, and the mating boss can be embedded in the limiting groove.

[0017] By adopting the above technical solution, the sealing film and glass slide are limited by the limiting groove on the cover and the mating boss on the channel seat, thereby facilitating the installation of the sealing film and glass slide.

[0018] Preferably, the cavity opening includes a connecting portion, a liquid guiding portion, and a reaction portion. Two connecting portions and two liquid guiding portions are provided. The two connecting portions are located at both ends of the cavity opening. The first sample port and the second sample port are respectively located in the two connecting portions. Two liquid guiding portions are provided. The width of the connecting portion is smaller than the width of the reaction portion. The reaction portion is connected to the connecting portion through the liquid guiding portion.

[0019] By adopting the above technical solution, when the reagent enters the reaction chamber through the first sample port, it first passes through the connecting part and then along the liquid guiding part to the reaction part at the position of the tissue sample. Since the width of the connecting part is smaller than the width of the reaction part, the liquid guiding part is inclined towards the reaction part on both sides, so that the reagent is evenly diffused into the entire reaction chamber along the liquid guiding part, thereby making the tissue sample react fully.

[0020] Preferably, two buckle assemblies are provided, and the two buckle assemblies are provided on both sides of the cover; the buckle assembly includes a connecting block and a snap-fit ​​block, one end of the connecting block is rotatably connected to the side wall of the cover, and the other end of the connecting block is rotatably connected to the snap-fit ​​block, and the snap-fit ​​block can snap onto the surface of the channel seat away from the cover.

[0021] By adopting the above technical solution, the connecting blocks on both sides of the cover and the snap-fit ​​blocks facilitate the assembly and disassembly of the cover and the channel seat. At the same time, the snap-fit ​​blocks are fixed on the surface of the channel seat away from the cover, thereby reducing contact with the snap-fit ​​blocks during observation or imaging scanning, thus improving the stability of the cover and the channel seat.

[0022] Preferably, the side wall of the cover is provided with an installation notch, the connecting block is connected to the installation notch via a rotating shaft, the side wall of the channel seat is provided with a limiting notch corresponding to the installation notch, and the connecting block can be embedded in the limiting notch.

[0023] By adopting the above technical solution, the connecting block is limited by the limiting notch, thereby improving the stability of the snap-fit ​​assembly.

[0024] Preferably, the sealing sheet is a silicone sheet.

[0025] By adopting the above technical solution, the silica gel sheet does not react with the reagent, and at the same time, it can form a closed reaction chamber. In addition, it can also play a buffering role to avoid damage to the glass slide.

[0026] Preferably, it also includes a soft pad placed between the glass slide and the cover.

[0027] By adopting the above technical solution, the soft gasket placed between the slide and the cover can further protect the slide.

[0028] Preferably, the side wall of the channel seat is provided with a magnetic suction element.

[0029] By adopting the above technical solution, the magnetic components on the channel seat can be quickly connected to the external injection pump system.

[0030] In summary, this application includes at least one of the following beneficial technical effects: First, a sealing film is placed on the surface of the channel seat. By adjusting the position of the sealing film, the first sample port and the second sample port are located at opposite ends of the cavity opening. Then, a glass slide is placed on the sealing film to form a reaction cavity. The position of the glass slide is adjusted horizontally so that the tissue sample on the glass slide is located inside the cavity opening. Then, a cover is placed on the glass slide, and the cover is fixed to the channel seat by a snap-fit ​​assembly. The cover is connected to a multi-port selector valve through a first pair of interfaces, and to an injection pump through a second pair of interfaces. Thus, the reagent in the reagent tube is injected into the reaction cavity for tissue sample incubation and staining. Therefore, contamination during the tissue sample incubation and staining process is reduced, and the accuracy of sequencing sample preparation is improved. Attached Figure Description

[0031] Figure 1 This is a front view of the chip adapter in an embodiment of this application.

[0032] Figure 2 This is an exploded structural diagram of the channel seat, sealing film, cover glass, and cover plate in the embodiments of this application.

[0033] Figure 3 yes Figure 1 Sectional view of AA.

[0034] Figure 4 This is a schematic diagram of the structure of sealing sheets of different capacities in the embodiments of this application.

[0035] Figure 5 This is an exploded structural diagram of the channel seat, sealing film, glass slide, soft gasket, and cover plate in the embodiments of this application.

[0036] Figure 6 This is a schematic diagram of the structure of the carrier adapter after assembly in an embodiment of this application.

[0037] Explanation of reference numerals in the attached drawings: 1. Glass slide; 2. Channel seat; 21. First channel; 22. Second channel; 23. First interface; 24. Second interface; 25. First sample port; 26. Second sample port; 27. Magnetic suction element; 28. Mating boss; 29. ​​Limiting notch; 3. Sealing soft sheet; 31. Cavity opening; 311. Connecting part; 312. Liquid guiding part; 313. Reaction part; 4. Cover; 41. Transparent window; 42. Limiting groove; 43. Installation notch; 5. Snap-fit ​​assembly; 51. Connecting block; 52. Snap-fit ​​block; 521. Snap-fit ​​part; 522. Moving part; 6. Soft gasket. Detailed Implementation

[0038] The following is in conjunction with the appendix Figure 1-6 This application will be described in further detail.

[0039] This application discloses a chip adapter for an in situ RNA sequencer. (Refer to...) Figure 1 and Figure 2 The slide adapter is used to mount a glass slide 1 containing a tissue sample, and a reaction chamber is formed between the slide adapter and the glass slide 1. The reaction chamber is connected to the injection pump system via the slide adapter, thereby injecting reagents into the reaction chamber for incubation and staining of the tissue sample.

[0040] Reference Figure 2 and Figure 3 Specifically, the carrier adapter includes a channel seat 2, a sealing film 3, a cover 4, and a snap-fit ​​assembly 5. The channel seat 2 is rectangular in shape. The channel seat 2 has a first channel 21 and a second channel 22 inside. One surface of the channel seat 2 has a first pair of interfaces 23 and a second pair of interfaces 24. One end of the first channel 21 communicates with the first pair of interfaces 23, and one end of the second channel 22 communicates with the second pair of interfaces 24. In this embodiment, the first pair of interfaces 23 and the second pair of interfaces 24 are located on one side wall of the channel seat 2. The other surface of the channel seat 2 has a first sample port 25 and a second sample port 26. The other end of the first channel 21 communicates with the first sample port 25, and the other end of the second channel 22 communicates with the second sample port 26. In this embodiment, the first sample port 25 and the second sample port 26 are located on the upper surface of the channel seat 2.

[0041] Reference Figure 2 The sealing sheet 3 has a cavity opening 31 located in the middle of the sealing sheet 3. The sealing sheet 3 is placed on the surface of the channel seat 2. The first sample port 25 and the second sample port 26 are distributed at both ends of the cavity opening 31. The glass slide 1 covers the upper surface of the sealing sheet 3. The cavity opening 31 of the sealing sheet 3, the surface of the channel seat 2, and the glass slide 1 form a reaction chamber. In this embodiment, the sealing sheet 3 is made of silicone. The silicone sheet does not react with the reagent and can form a sealed reaction chamber. In addition, it can also act as a buffer to prevent the glass slide 1 from being damaged by pressure.

[0042] The cover 4 covers the surface of the glass slide 1, and the snap-fit ​​assembly is connected to the cover 4 and can be snapped onto the channel seat 2. The side wall of the channel seat 2 is provided with a magnetic suction member 27, which allows for quick docking and connection with the external injection pump system.

[0043] Reference Figure 2Furthermore, to facilitate observation of the reagents within the reaction chamber and direct scanning imaging of the reacted tissue sample, a transparent window 41 is provided on the cover 4 corresponding to the cavity opening 31. To accelerate the reaction between the tissue sample and the reagents, the reaction chamber needs to be heated, typically to a temperature of 20-50 degrees Celsius, depending on the reagent type. The transparent window 41 allows for rapid heat transfer into the reaction chamber. The thickness of the transparent window 41 is less than the thickness of the cover 4. The periphery of the cover 4 is tilted towards the transparent window 41, creating a larger observation angle, thus facilitating observation of the reagent injection within the reaction chamber and enabling convenient scanning imaging.

[0044] Reference Figure 2 Furthermore, a limiting groove 42 is provided on the surface of the cover 4 facing the channel seat 2, and a mating boss 28 is raised on the surface of the channel seat 2. The sealing film 3 is placed on the mating boss 28, and the first sample port 25 and the second sample port 26 are spaced apart along the length direction on the mating boss 28. The mating boss 28 can be embedded in the limiting groove 42. The limiting groove 42 on the cover 4 and the mating boss 28 on the channel seat 2 limit the sealing film 3 and the glass slide 1, thereby facilitating the installation of the sealing film 3 and the glass slide 1, and positioning the first sample port 25 and the second sample port 26, so that the first sample port 25 and the second sample port 26 are located in the middle position of the cavity opening 31.

[0045] Reference Figure 4 The cavity opening 31 includes a connecting portion 311, a liquid guiding portion 312, and a reaction portion 313. Two connecting portions 311 and two liquid guiding portions 312 are provided. The two connecting portions 311 are located at opposite ends of the cavity opening 31. The first sample port 25 and the second sample port 26 are respectively located within the two connecting portions 311. Two liquid guiding portions 312 are provided. The width of the connecting portion 311 is smaller than the width of the reaction portion 313. The reaction portion 313 is connected to the connecting portion 311 through the liquid guiding portion 312. When the reagent enters the reaction cavity through the first sample port 25, it first passes through the connecting portion 311 and then along the liquid guiding portion 312 to the reaction portion 313. Because the width of the connecting portion 311 is smaller than the width of the reaction portion 313, the sides of the liquid guiding portion 312 are inclined towards the reaction portion 313, thereby allowing the reagent to diffuse evenly throughout the entire reaction cavity along the liquid guiding portion 312, ensuring a complete reaction of the tissue sample. Furthermore, by adjusting the lengths of the connecting portion 311 and the reaction portion 313, the capacity of the reaction chamber can be easily adjusted to accommodate different tissue sample capacities. Simultaneously, the capacity of the reaction chamber can also be adjusted by adjusting the thickness of the sealing film 3.

[0046] Reference Figure 5 and Figure 6Two latching components 5 are provided, located on both sides of the cover 4. Each latching component 5 includes a connecting block 51 and a latching block 52. One end of the connecting block 51 is rotatably connected to the side wall of the cover 4, and the other end of the connecting block 51 is rotatably connected to the latching block 52. The latching block 52 can latch onto the surface of the channel seat 2 away from the cover 4. The connecting blocks 51 and the latching blocks 52 on both sides of the cover 4 facilitate the assembly and disassembly of the cover 4 and the channel seat 2. At the same time, the latching blocks 52 are fixed on the surface of the channel seat 2 away from the cover 4, thereby reducing contact with the latching blocks 52 during observation or imaging scanning, thus improving the stability of the cover 4 and the channel seat 2.

[0047] Furthermore, the latching block 52 includes a latching part 521 and a toggle part 522. The toggle part 522 is integrally formed on the side of the latching part 521 away from the rotational connection. The latching part 521 can abut against the surface of the channel seat 2. The toggle part 522 facilitates the latching part 521 to latch and unlock the latching state.

[0048] The cover 4 has an installation notch 43 on its side wall. The connecting block 51 is connected to the installation notch 43 via a pivot. The channel seat 2 has a limiting notch 29 on its side wall that corresponds to the installation notch 43. The connecting block 51 can be embedded in the limiting notch 29. The connecting block 51 is limited by the limiting notch 29, thereby improving the stability of the buckle assembly 5.

[0049] Furthermore, a soft pad 6 is included, which is placed between the glass slide 1 and the cover plate 4. The glass slide 1 includes a slide and a cover plate. Depending on the testing scenario, either a slide or a cover plate may be used. The cover plate is about 0.15 mm thick, and the slide is about 1.1 mm thick. Therefore, when using a cover plate, a soft pad 6 needs to be placed on the upper surface of the cover plate. In this embodiment, the soft pad 6 is a silicone sheet.

[0050] The implementation principle of a chip adapter for an in situ RNA sequencer according to an embodiment of this application is as follows: First, a sealing film 3 is placed on the surface of the channel seat 2. By adjusting the position of the sealing film 3, the first sample port 25 and the second sample port 26 are located at both ends of the cavity port 31. Then, a glass slide 1 is placed on the sealing film 3, and the position of the glass slide 1 is adjusted horizontally so that the tissue sample on the glass slide 1 is located inside the cavity port 31, forming a reaction chamber for the tissue sample. Then, a cover 4 is placed on the glass slide 1, and the cover 4 is fixed to the channel seat 2 by means of a snap-fit ​​assembly 5. The reagent is injected into the reaction chamber for tissue sample incubation and staining by connecting to a syringe pump through the first pair of interfaces 23 and the second pair of interfaces 24. Therefore, the contamination during the tissue sample incubation and staining process is reduced, and the accuracy of sequencing sample preparation is improved.

[0051] 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 chip adapter for an in situ RNA sequencer, characterized in that, include: A channel seat (2) is provided inside, which has a first channel (21) and a second channel (22); one surface of the channel seat (2) is provided with a first pair of interfaces (23) and a second pair of interfaces (24), one end of the first channel (21) is connected to the first pair of interfaces (23), and one end of the second channel (22) is connected to the second pair of interfaces (24); the other surface of the channel seat (2) is provided with a first sample port (25) and a second sample port (26), the other end of the first channel (21) is connected to the first sample port (25), and the other end of the second channel (22) is connected to the second sample port (26); A sealing sheet (3) is provided with a cavity opening (31). The sealing sheet (3) is placed on the surface of the channel seat (2). The first sample opening (25) and the second sample opening (26) are distributed at both ends of the cavity opening (31). The sealing sheet (3), the channel seat (2), and the glass slide (1) form a reaction cavity. The cavity opening (31) includes a connecting part (311), a liquid guiding part (312), and a reaction part (313). Two connecting parts (311) and two liquid guiding parts (312) are provided. The two connecting parts (311) are located at both ends of the cavity opening (31). The first sample port (25) and the second sample port (26) are respectively located in the two connecting parts (311). Two liquid guiding parts (312) are provided. The width of the connecting part (311) is smaller than the width of the reaction part (313). The reaction part (313) is connected to the connecting part (311) through the liquid guiding part (312). Cover (4), the cover (4) is used to cover the glass slide (1) on the surface of the sealing film (3); The snap-fit ​​assembly (5) is connected to the cover (4) and can be snapped onto the channel seat (2), and the channel seat (2) is provided with a magnetic suction element embedded in its side wall.

2. The chip adapter for the in situ RNA sequencer according to claim 1, characterized in that: The cover (4) is provided with a transparent window (41) at the position of the cavity opening (31), and the thickness of the transparent window (41) is less than the thickness of the cover (4).

3. The chip adapter for the in situ RNA sequencer according to claim 2, characterized in that: The cover (4) is inclined around the periphery of the transparent window (41).

4. The chip adapter for the in situ RNA sequencer according to claim 1, characterized in that: The cover (4) has a limiting groove (42) on its surface facing the channel seat (2). The channel seat (2) has a protruding mating boss (28) on its surface. The sealing sheet (3) is placed on the mating boss (28). The first sample port (25) and the second sample port (26) are distributed at intervals along the length direction on the mating boss (28). The mating boss (28) can be embedded in the limiting groove (42).

5. The chip adapter for the in situ RNA sequencer according to claim 1, characterized in that: Two buckle assemblies (5) are provided, and the two buckle assemblies (5) are provided on both sides of the cover (4); the buckle assembly (5) includes a connecting block (51) and a snap-fit ​​block (52). One end of the connecting block (51) is rotatably connected to the side wall of the cover (4), and the other end of the connecting block (51) is rotatably connected to the snap-fit ​​block (52). The snap-fit ​​block (52) can snap onto the surface of the channel seat (2) away from the cover (4).

6. The chip adapter for the in situ RNA sequencer according to claim 5, characterized in that: The cover (4) has an installation notch (43) on its side wall. The connecting block (51) is connected to the installation notch (43) by a pivot. The channel seat (2) has a limiting notch (29) on its side wall that corresponds to the installation notch (43). The connecting block (51) can be embedded in the limiting notch (29).

7. The chip adapter for the in situ RNA sequencer according to claim 1, characterized in that: The sealing sheet (3) is a silicone sheet.

8. The chip adapter for the in situ RNA sequencer according to claim 1, characterized in that: It also includes a soft pad (6) placed between the glass slide (1) and the cover (4).

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