Dissociation module and dissociation device

Abstract

The application discloses a dissociation module and a dissociation device. The dissociation module comprises a plurality of barrels for storing liquid materials and a mounting seat for connecting the plurality of barrels into one body. The plurality of barrels comprise a dissociation barrel, a liquid forming barrel and a liquid storage barrel. The dissociation barrel movably has a cutter. The cutter is used for crushing tissues in the dissociation barrel in the movement stroke. The liquid forming barrel is communicated with the dissociation barrel and is used for storing single-cell suspension obtained by dissociation. The liquid storage barrel is used for storing dissociation additives and is communicated with the dissociation barrel and / or the liquid forming barrel. The dissociation module is used for being connected with a power assembly in the dissociation device, so as to drive the cutter to move and drive the liquid materials to flow along the communication between the two barrels by the power assembly. The application can realize the orderly, accurate and efficient performance of each process in the dissociation module, help to reduce the operation dependence on the experimental personnel and reduce the error rate of the dissociation process.

Description

Technical Field

[0001] This invention relates to the field of tissue dissociation technology, specifically to a dissociation module and dissociation device. Background Technology

[0002] In biological research, technologies such as single-cell sequencing require the processing and analysis of individual cells. To obtain single cells for research, current technologies generally require first breaking down and dispersing bulk animal and plant tissues, such as experimental mouse heart tissue or artificially cultured human kidney tissue, into single-cell suspensions.

[0003] Currently, the technology for dissociating tissues into single-cell suspensions is relatively mature, but the dissociation procedure is quite complex. It requires experimenters to manually handle various dissociation tubes and other related equipment, and then manually add liquids one by one. This requires high skill from the experimenters and has a low margin for error. Summary of the Invention

[0004] The main objective of this invention is to propose a dissociation module and dissociation device, which aims to solve the problems of traditional tissue dissociation equipment relying on manual operation by laboratory personnel, resulting in complex operation and low error tolerance.

[0005] To achieve the above objectives, the present invention proposes a dissociation module for installation in a dissociation device. The dissociation module includes multiple cylinders for storing liquid and a mounting base connecting the multiple cylinders into a single unit. The multiple cylinders include:

[0006] The dissociation cylinder is equipped with a movably mounted cutter, which is used to break up the tissue inside the dissociation cylinder during its moving stroke;

[0007] A liquid-forming cylinder, connected to the dissociation cylinder, is used to store the single-cell suspension obtained from dissociation; and...

[0008] A storage tank for storing dissociation additives and connected to the dissociation tank and / or the liquid-forming tank;

[0009] The dissociation module is connected to the power component in the dissociation device to drive the cutter movement and drive the liquid material to flow along the connection between the two cylinders.

[0010] Optionally, the two connected cylinders are a first cylinder and a second cylinder, respectively;

[0011] The dissociation module further includes an installation cavity for the removable installation of a pressure regulating component in the power assembly. The installation cavity is connected to the first cylinder to adjust the pressure inside the first cylinder, thereby controlling the liquid to be drawn from the second cylinder into the first cylinder or discharged from the first cylinder into the second cylinder.

[0012] Optionally, the plurality of cylinders includes a third cylinder, which includes a body and a bottom plate. The body has a cavity extending through it along its axial direction and forms two openings. The bottom plate is movably installed in the cavity along the axial direction of the body and is in a movable sealing fit with the inner wall of the cavity, so that under the drive of the telescopic component in the power assembly, the bottom plate moves from one opening to the other opening and pushes out the liquid in the third cylinder.

[0013] Optionally, the cutting tool includes:

[0014] The first cutter body includes a shaft rotatably disposed about the axis of the dissociation cylinder and a rib protruding from the side wall of the shaft. The shaft is connected to a rotary drive component in the power assembly, and the rib has a downward-facing first cutting face.

[0015] The second blade body protrudes from the periphery of the shaft body below the rib, and the second blade body has a second blade surface facing upward.

[0016] The second cutting surface is spaced apart from the first cutting surface, and the space defines a grinding area. During the rotation stroke of the first cutting body, the first cutting surface and the second cutting surface rotate relative to each other, and the tissue passing through the grinding area is ground.

[0017] Optionally, the rib extends spirally along the circumferential and axial directions of the shaft.

[0018] The helix angle of the raised rib gradually decreases from top to bottom; and / or,

[0019] The height of the protruding ribs from the shaft body gradually decreases from bottom to top.

[0020] Optionally, the second cutter body is provided with a plurality of cutters arranged sequentially along the circumference of the shaft; and / or,

[0021] The second cutting face extends circumferentially and upward from the bottom wall of the dissociation cylinder towards the shaft body. The second cutting body also has a third cutting face connecting the top end of the second cutting face and the bottom wall of the dissociation cylinder. The inclination angle between the second cutting face and the bottom wall of the dissociation cylinder is not less than the inclination angle between the third cutting face and the bottom wall of the dissociation cylinder.

[0022] Optionally, the upper end of the shaft is provided with a groove facing downwards, and a through hole is provided in the groove near the bottom wall of the dissociation cylinder, and the groove is at least in communication with the liquid forming cylinder.

[0023] Optionally, the cylindrical body has a first cylindrical section extending upward from the mounting base. This first cylindrical section is flared from bottom to top and is made of an elastic material, so that when the cover plate in the dissociation device closes the openings of each cylindrical body, the first cylindrical section deforms and adheres to the cover plate; and / or,

[0024] The liquid forming cylinder is equipped with a filter screen, which is used to filter the liquid entering the liquid forming cylinder.

[0025] Furthermore, to achieve the above objectives, the present invention also provides a dissociation device, comprising a main body, a power component disposed on the main body, and a dissociation module, wherein the dissociation module is detachably connected to the main body and the power component, and the dissociation module includes a plurality of cylinders for storing liquid and a mounting base for connecting the plurality of cylinders into one unit, wherein the plurality of cylinders include:

[0026] The dissociation cylinder is equipped with a movably mounted cutter, which is used to break up the tissue inside the dissociation cylinder during its moving stroke;

[0027] A liquid-forming cylinder, connected to the dissociation cylinder, is used to store the single-cell suspension obtained from dissociation; and...

[0028] A storage tank for storing dissociation additives and connected to the dissociation tank and / or the liquid-forming tank;

[0029] The dissociation module is connected to the power component in the dissociation device to drive the cutter movement and drive the liquid material to flow along the connection between the two cylinders.

[0030] Optionally, the power assembly includes a pressure regulating component, and / or a telescopic component, and / or a rotary drive component; and / or,

[0031] The main body includes a base and a cover plate. The base is provided with an installation groove for installing the dissociation module. The cover plate is movably installed on the cover plate so as to simultaneously open and close the groove of the installation groove and the opening of each of the cylinders.

[0032] In the technical solution provided by this invention, each cylinder in the dissociation module is used to store liquid materials, which can directly participate in various tissue dissociation processes or be reserved for later use. The liquid materials can vary depending on the function of the cylinder. When the cylinder is a dissociation cylinder, the liquid material is tissue with fragmentation. When the cylinder is a liquid-forming cylinder, the liquid material is an intermediate or final product of the dissociation operation, a single-cell suspension. When the cylinder is a liquid storage cylinder, the liquid material is an additive required in the dissociation process, such as lysis buffer, lysinusoidal lysate, and termination solution. The mounting base connects each cylinder into a single unit, forming a modular design, which helps to centrally group the various dissociation-related equipment and liquid materials, avoiding omissions in the process. After the dissociation module is installed in the dissociation device, with the help of the power component in the dissociation device, the various processes in the dissociation module can be carried out in an orderly, accurate, and efficient manner, which helps to reduce the dependence on the operation of experimental personnel and reduce the error rate of the dissociation process. Attached Figure Description

[0033] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0034] Figure 1 A perspective view of an embodiment of the dissociation module provided by the present invention;

[0035] Figure 2 for Figure 1 Top view of the dissociation module;

[0036] Figure 3 for Figure 1 A bottom view of the dissociation module;

[0037] Figure 4 for Figure 1 Schematic diagram of the longitudinal section of the dissociation module;

[0038] Figure 5 for Figure 4 Enlarged structural diagram at point A;

[0039] Figure 6 for Figure 1 A schematic diagram of the structure after the first cutter body has been removed;

[0040] Figure 7 for Figure 6 Enlarged structural diagram at point B;

[0041] Figure 8 for Figure 1 A three-dimensional schematic diagram of the first blade.

[0042] Explanation of icon numbers:

[0043]

[0044]

[0045] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0046] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0047] It should be noted that if the embodiments of the present invention involve directional indicators (such as up, down, left, right, front, back, etc.), the directional indicators are only used to explain the relative positional relationship and movement of the components in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indicators will also change accordingly.

[0048] Furthermore, if the embodiments of this invention involve descriptions such as "first" or "second," these descriptions are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include at least one of those features. Additionally, the meaning of "and / or" throughout the text includes three parallel solutions; for example, "A and / or B" includes solution A, solution B, or a solution where both A and B are satisfied simultaneously. Furthermore, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or impossible to implement, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection claimed by this invention.

[0049] In biological research, technologies such as single-cell sequencing require the processing and analysis of individual cells. To obtain single cells for research, current technologies generally require first breaking down and dispersing bulk animal and plant tissues, such as experimental mouse heart tissue or artificially cultured human kidney tissue, into single-cell suspensions.

[0050] Currently, the technology for dissociating tissues into single-cell suspensions is relatively mature, but the dissociation procedure is quite complex. It requires experimenters to manually handle various dissociation tubes and other related equipment, and then manually add liquids one by one. This requires high skill from the experimenters and has a low margin for error.

[0051] In view of the above, the present invention provides a dissociation device.

[0052] In this design, the dissociation device includes a main body, a power component, and a dissociation module 1. The dissociation module 1 includes multiple cylinders 100 for storing liquid material and a mounting base 200 connecting the multiple cylinders 100 into a single unit. Each cylinder 100 includes a dissociation cylinder 110, a liquid-forming cylinder 120, and a liquid-retaining cylinder 130. A cutter 300 is movably mounted on the dissociation cylinder 110, which breaks down the tissue within the dissociation cylinder 110 during its stroke. The liquid-forming cylinder 120 communicates with the dissociation cylinder 110 and stores the dissociated single-cell suspension. The liquid-retaining cylinder 130 stores dissociation additives and communicates with the dissociation cylinder 110 and / or the liquid-forming cylinder 120. The dissociation module 1 is connected to the power component of the dissociation device to drive the cutter 300 and drive the liquid material to flow along the connection between two cylinders 100.

[0053] In the technical solution provided by this invention, each cylinder 100 in the dissociation module 1 is used to store liquid materials, which can directly participate in various tissue dissociation processes or be reserved for later use. The liquid materials can vary depending on the function of the cylinder 100. When the cylinder 100 is a dissociation cylinder 110, the liquid material is tissue with broken tissue. When the cylinder 100 is a liquid-forming cylinder 120, the liquid material is an intermediate or final product of the dissociation operation, such as a single-cell suspension. When the cylinder 100 is a liquid storage cylinder 130, the liquid material is an additive used in the dissociation process, such as lysis buffer, red blood cell lysis buffer, and termination solution. The mounting base 200 connects each cylinder 100 into a single unit, forming a modular design. This helps to centrally group the various dissociation-related equipment and liquid materials, avoiding omissions in the process. After the dissociation module 1 is installed in the dissociation device, with the help of the power component in the dissociation device, the various processes in the dissociation module 1 can be carried out in an orderly, accurate, and efficient manner, which helps to reduce the dependence on the operation of experimental personnel and reduce the error rate of the dissociation process.

[0054] Specifically, the main body may include a casing, which is used to mount the power assembly and the dissociation module 1. The main body generally integrates a control module, which may, but is not limited to, consist of, for example, a power supply structure, a controller, and various sensors.

[0055] The power assembly is electrically connected to the controller. The power assembly can be configured differently depending on the different requirements of the dissociation module 1. Specifically, for example, the power assembly includes a voltage regulating component, and / or a telescopic component, and / or a rotary drive component.

[0056] The dissociation module 1 is detachably connected to the body of the dissociation device. When installed on the body, the dissociation module 1 can be simultaneously connected to the power component. When the dissociation module 1 is removed from the body, the dissociation module 1 can be simultaneously disconnected from the power component.

[0057] The dissociation module 1 is used to dissociate plant and animal tissues to obtain the desired single-cell suspension. In one embodiment, the dissociation module 1 itself or the body further includes components for cleaning, disinfection, and sterilization, which can clean, disinfect, and sterilize each cylinder 100 in the dissociation device after each dissociation operation, ensuring that each cylinder 100 meets the purification requirements for the next tissue dissociation. This allows the dissociation module 1 to be reused multiple times.

[0058] In order to reduce the need for additional components for cleaning, disinfection, and sterilization, and to ensure that each dissociation operation is clean and free from harmful effects, in this embodiment, the dissociation module 1 is a single-use structure, so that each tissue dissociation operation uses a brand new dissociation module 1.

[0059] Based on this, a positioning structure can be provided between the body and the dissociation module 1, and / or between the power component and the dissociation module 1, so as to achieve quick and accurate installation of each dissociation module 1 on the body and / or at the connection point with the power component.

[0060] Taking the connection between the power component and the dissociation module 1 as an example, there are various solutions for the positioning structure, such as:

[0061] The positioning structure can be a positioning protrusion and a positioning recess. One of the power component and the disengagement module 1 is provided with a positioning protrusion, and the other with a positioning recess. The positioning connection between the power component and the disengagement module 1 is achieved through the matching of the positioning protrusion and the positioning recess. The positioning protrusion is, for example, a snap-fit, and the positioning recess is, for example, a snap-fit ​​groove or a snap-fit ​​hole.

[0062] The positioning structure can also be a magnetic component and a magnetic mating component. One of the power assembly and the dissociation module 1 is provided with a magnetic component, and the other with a magnetic mating component. Positioning connection between the power assembly and the dissociation module 1 is achieved through magnetic attraction between the magnetic component and the magnetic mating component. The magnetic component is, for example, a magnetic sheet, and the magnetic mating component is, for example, a sheet-like structure made of materials such as iron, cobalt, or nickel.

[0063] Based on the above, by mounting the power assembly on the body and keeping it separate from the dissociation module 1 when not in operation, the independence of the dissociation module 1 can be ensured, while reducing its economic cost, simplifying its structure, and lightening its weight. The power assembly can be adapted to different dissociation modules 1 and can be reused.

[0064] Since the present invention mainly improves the dissociation module 1, the following will be combined with the appendix. Figures 1 to 8 The specific embodiments of the dissociation module 1 are described, mainly focusing on the structure of the dissociation module 1.

[0065] Please see Figures 1 to 3 As described above, the dissociation module 1 includes a plurality of cylinders 100 for storing liquid and a mounting base 200 for connecting the plurality of cylinders 100 into a single unit.

[0066] The specific forms of the plurality of cylinders 100, such as size, shape, and material, can be identical or at least partially different. When the plurality of cylinders 100 have the same specific form, the dissociation module 1 further includes an identifier to distinguish each cylinder 100. The identifier is disposed on at least a portion of the cylinders 100 to differentiate them. The identifier can be various patterns, characters, color blocks, embossed structures, etc. When the plurality of cylinders 100 have at least partially different specific forms, the different cylinders 100 can have, for example, different volumes, shapes, colors, and materials. This not only helps to distinguish the individual cylinders 100 but also improves the compatibility between the cylinder 100 and the liquid it stores. For example, when the liquid stored in the cylinder 100 needs to be protected from light, the cylinder 100 can be configured as a dark-colored, opaque structure; when the liquid stored in the cylinder 100 has high temperature requirements, the cylinder 100 can be configured as having a heat insulation structure, or a temperature regulating component can be provided on the outside or inside of the cylinder 100, etc.

[0067] The mounting base 200 connects the plurality of cylinders 100 into a single unit. Specifically, the mounting base 200 may be integrally formed with the plurality of cylinders 100, or the mounting base 200 may be separately formed from the plurality of cylinders 100, and the two may be tightly connected by means of fastening, screwing, bonding or adsorption.

[0068] The mounting base 200 can be configured as one, and is generally plate-shaped or block-shaped; the mounting base 200 can also be configured as multiple, and is disposed between every two adjacent cylinders 100, so that the multiple cylinders 100 can be connected and fixed in pairs.

[0069] The end faces of the openings of the plurality of cylinders 100 are generally kept on the same plane. The mounting base 200 can be disposed at any suitable position along the length of the cylinder 100, for example, disposed sufficiently close to the opening of each cylinder 100, disposed sufficiently far from the opening of each cylinder 100, or disposed in between. In this embodiment, the mounting base 200 is disposed relatively closer to the opening of each cylinder 100.

[0070] The dissociation cylinder 110 is equipped with a cutter 300, which is movably disposed relative to the dissociation cylinder 110. The specific movement of the cutter 300 is not limited; it can be a reciprocating translation, relative rotation, or a combination of translation and rotation in any direction. In specific applications, depending on different tissues and requirements, the cutter 300 can be configured to break down tissues through methods such as compression, grinding, puncture, or cutting. Therefore, the specific structure of the cutter 300 can be configured differently depending on the different breaking principles.

[0071] The liquid-forming cylinder 120 is connected to the dissociation cylinder 110 and is used to store the single-cell suspension obtained by dissociation. It should be noted that the above does not constitute a limitation on the type of liquid material stored in the liquid-forming cylinder 120 during the dissociation process. It can be understood that the liquid-forming cylinder 120 is ultimately used to store the single-cell suspension, but it can also be used to store the required intermediate products during the entire dissociation process.

[0072] The liquid-forming cylinder 120 and the mounting base 200 are detachably connected via a connecting structure 600, so that the liquid-forming cylinder 120 and the mounting base 200 remain connected during the dissociation process. This connection can be fixed or movable. After dissociation is completed and a single-cell suspension is obtained, the liquid-forming cylinder 120 can be removed from the mounting base 200 by a drive mechanism on the machine or manually by the experimenter, so as to facilitate the subsequent transfer, processing and storage of the single-cell suspension in the liquid-forming cylinder 120.

[0073] There are various specific designs for the connection structure 600, such as a screw connection between an internal thread structure and an external thread structure, a snap-fit ​​fixing between a snap fastener and a snap groove, a magnetic adsorption between a magnetic component and a magnetic mating component, or an adhesive fixing between an adhesive component and an adhesive mating component, etc., without any limitation.

[0074] The liquid forming cylinder 120 can be specifically configured as test tubes of various specifications and styles, and can be standard or non-standard parts.

[0075] The storage tank 130 is used to store the additives required during the dissociation process. It should be noted that any one of the dissociation tank 110, the liquid-forming tank 120, and the storage tank 130 can be one or more. For example, there can be three storage tanks 130, namely a pyrolysis liquid tank 131, a red pyrolysis liquid tank 133, and a termination liquid tank 132. As their names suggest, the pyrolysis liquid tank 131 is used to store the pyrolysis liquid, the red pyrolysis liquid tank 133 is used to store the red pyrolysis liquid, and the termination liquid tank 132 is used to store the termination liquid.

[0076] Furthermore, the arrangement of the multiple cylinders 100, such as the dissociation cylinder 110, the liquid-forming cylinder 120, and the liquid storage cylinder 130, is not limited. For example, depending on actual needs, two cylinders 100 that are connected can be arranged close together, while two cylinders 100 that are not connected can be arranged apart. Specifically, being connected can be manifested as a connecting pipe being provided between two cylinders 100.

[0077] In the above embodiments, the power assembly is at least capable of driving the cutter 300 to move and the liquid to flow through the connection between the two cylinders 100, so as to transfer the liquid from one cylinder 100 to another. For ease of understanding, in the following embodiments, any two cylinders 100 that are connected are defined as the first cylinder 101 and the second cylinder 102. The first cylinder 101 and the second cylinder 102 can be connected at any position. The dissociation module 1 also includes a mounting cavity 140, which is used for the detachable installation of the pressure regulating component in the power assembly. The mounting cavity 140 is connected to the first cylinder 101 to adjust the pressure inside the first cylinder 101, so as to control the liquid to be drawn from the second cylinder 102 into the first cylinder 101 or discharged from the first cylinder 101 into the second cylinder 102.

[0078] The mounting cavity 140 may be defined by one of the plurality of cylinders 100, or it may be defined by an additional shell structure in addition to the plurality of cylinders 100. The mounting cavity 140 may be used for the fixed installation of the power component after the dissociation module 1 is installed on the body of the dissociation device; or the mounting cavity 140 may serve as a transition channel for the flow of, for example, external blowing air or internal extraction air.

[0079] The pressure regulating component, such as a pump, a blower, a vacuum device, or a temperature regulating device, can create a pressure difference between the first cylinder 101 and the second cylinder 102, thereby enabling the transfer of liquid material between the first cylinder 101 and the second cylinder 102.

[0080] In one embodiment, the plurality of cylinders 100 includes a third cylinder 103, that is, any one of the plurality of cylinders 100 can be defined as the third cylinder 103. The third cylinder 103 includes a cylinder body 103a and a bottom plate 103b. The cylinder body 103a has a cavity extending through it along its axial direction and forms two openings. The bottom plate 103b is movably installed in the cavity along the axial direction of the cylinder body 103a and is in a movable sealing fit with the inner wall of the cavity, so that under the drive of the telescopic component in the power assembly, the bottom plate 103b translates from one opening to the other opening and pushes out the liquid in the third cylinder body 103a.

[0081] It can be understood that the base plate 103b is equivalent to a piston, enabling the intake and ejection of liquid material during its reciprocating movement. Based on this, the telescopic component can be, for example, a telescopic cylinder, or a component composed of a drive motor and a linear transmission assembly, such as a lead screw and nut mechanism, a gear and rack mechanism, a worm gear mechanism, or a linkage mechanism.

[0082] Based on any of the above embodiments, please refer to Figures 4 to 5 The cutting tool 300 includes a first cutting body 310 and a second cutting body 320. The first cutting body 310 includes a shaft 311 rotatably disposed about the axis of the dissociation cylinder 110 and a rib 312 protruding from the side wall of the shaft 311. The shaft 311 is connected to a rotary drive component in the power assembly. The rib 312 has a downward-facing first cutting surface 312a. The second cutting body 320 is disposed around the periphery of the shaft 311 and protrudes below the rib 312. The second cutting body 320 has an upward-facing second cutting surface 321. The second cutting surface 321 is spaced apart from the first cutting surface 312a, and the space defines a grinding area. During the rotation stroke of the first cutting body 310, the first cutting surface 312a and the second cutting surface 321 are driven to rotate relative to each other, and the tissue passing through the grinding area is ground.

[0083] The second blade 320 may be fixed relative to the dissociation cylinder 110, while the first blade 310 may be movable relative to the dissociation cylinder 110; or, both the first cylinder 101 and the second cylinder 102 may be movable relative to the dissociation cylinder 110, but there is a difference in linear velocity between them, which causes relative movement between the first blade surface 312a and the second blade surface 321. This relative movement causes the tissue in the grinding area to be subjected to forces in opposite directions, thereby achieving grinding.

[0084] In the first cutter body 310, the shape and size of the protruding rib 312 are not limited; the protruding rib 312 can be one or at least two. The protruding rib 312 can be integrally formed with the shaft body 311, or it can be separately formed from the shaft body 311.

[0085] Specifically, please refer to Figure 5 and Figure 8 In one embodiment, the rib 312 extends spirally along the circumferential and axial directions of the shaft 311. This configuration ensures that when the shaft 311 drives the rib 312 to rotate in the opposite direction of the spiral of the rib 312 (e.g., clockwise), the interaction between the first cutting surface 312a and the second cutting surface 321 on the rib 312 is enhanced. The first cutting surface 312a can press the tissue in the grinding area downwards, strengthening the grinding effect between the first cutting surface 312a and the second cutting surface 321. Conversely, when the shaft 311 drives the rib 312 to rotate in the same direction as the spiral of the rib 312 (e.g., counterclockwise), the interaction between the first cutting surface 312a and the second cutting surface 321 on the rib 312 is weakened. The first cutting surface 312a can scoop up the tissue in the grinding area, causing the tissue to tumble in the dissociation cylinder 110. This helps to change the grinding direction of the first cutting surface 312a and the second cutting surface 321 on the tissue, and helps the tissue to be ground quickly and efficiently.

[0086] Based on the above, in one embodiment, the helix angle of the rib 312 gradually decreases from top to bottom. This arrangement makes the upper half of the rib 312 steeper, which helps to make the tissue fragments scooped up slide down more quickly; at the same time, it makes the lower half of the rib 312 flatter, and the first cutting surface 312a also flatter, which can increase the facing area between the first cutting surface 312a and the second cutting surface 321, thereby expanding the range of the grinding area and helping to improve grinding efficiency.

[0087] And / or, based on the above, further, in one embodiment, the height of the protrusion rib 312 protruding from the shaft 311 gradually decreases from bottom to top. This arrangement results in a relatively larger gap between the outer end of the upper portion of the protrusion rib 312 (i.e., the free end of the protrusion rib 312 radially away from the shaft 311) and the inner wall of the dissociation cylinder 110, facilitating the downward fall of upwardly scraped tissue fragments through this gap; simultaneously, it results in a relatively smaller gap between the outer end of the lower portion of the protrusion rib 312 and the inner wall of the dissociation cylinder 110, allowing the area between the outer end of the protrusion rib 312 and the inner wall of the dissociation cylinder 110 to simultaneously serve as a grinding area, thus improving grinding efficiency.

[0088] Please see Figure 6 and Figure 7 In one embodiment, multiple second blades 320 are arranged sequentially along the circumference of the shaft 311, such that when the first blade 310 rotates clockwise as described above, it can cooperate with the second blade 320 during the full circumference rotation of the first blade 310 to continuously grind the tissue in the grinding area; and when the first blade 310 rotates counterclockwise as described above, it can continuously scoop up the tissue in the grinding area during the full circumference rotation of the first blade 310 to achieve sufficient material turning.

[0089] And / or, based on the above, further, in one embodiment, the second cutting surface 321 extends circumferentially and upwardly from the bottom wall of the dissociation cylinder 110 toward the shaft 311, and the second cutting body 320 also has a third cutting surface 322 connecting the top end of the second cutting surface 321 and the bottom wall of the dissociation cylinder 110, wherein the inclination angle between the second cutting surface 321 and the bottom wall of the dissociation cylinder 110 is not less than the inclination angle between the third cutting surface 322 and the bottom wall of the dissociation cylinder 110. When the first blade body 310 rotates clockwise, the second blade surface 321 faces the first blade surface 312a. The inclination angle between the second blade surface 321 and the bottom wall of the dissociation cylinder 110 is set to be relatively large, which can increase the degree of interference between the first blade surface 312a and the second blade surface 321, thereby enhancing the grinding effect. Conversely, when the first blade body 310 rotates counterclockwise, the third blade surface 322 faces the first blade surface 312a. The inclination angle between the second blade surface 321 and the bottom wall of the dissociation cylinder 110 is set to be relatively small, which can reduce the degree of interference between the first blade surface 312a and the third blade surface 322. Furthermore, the third blade surface 322 forms a longer mating surface, which helps the rib 312 to scoop up the material on the third blade surface 322, thereby enhancing the material turning effect.

[0090] As can be seen from the above, during the dissociation process, the first cutter body 310 can be set to alternate between forward and reverse rotation through the control module, which helps the tissue to be broken up more quickly and completely.

[0091] Since there is a linear velocity difference between the first cutter body 310 and the second cutter body 320, in order to ensure a smoother relative rotation process, in one embodiment, a relief groove 313 with the groove facing downward and extending through the rotation direction of the shaft body 311 can be provided on the shaft body 311 or the rib 312. A protrusion 323 is provided on the second cutter body 320. The cooperation between the protrusion 323 and the relief groove 313 can guide and correct the relative rotation between the first cutter body 310 and the second cutter body 320.

[0092] In one embodiment, the upper end of the shaft 311 has a downward-facing groove 311a, and a through hole 311b is provided in the groove 311a near the bottom wall of the dissociation cylinder 110. The groove 311a communicates with at least the liquid-forming cylinder 120. The broken tissue / homogeneous solution formed by the mutual grinding of the first blade 310 and the second blade 320 in the dissociation cylinder 110 can enter the groove 311a through the through hole 311b. The dissociation module 1 may also include a conduit 500, one end of which communicates with, for example, the liquid-forming cylinder 120, and the other end of which extends downward into the groove 311a, so that the broken tissue / homogeneous solution formed by the mutual grinding of the first blade 310 and the second blade 320 in the dissociation cylinder 110 is extracted into the liquid-forming cylinder 120.

[0093] Furthermore, in one embodiment, the cylindrical body 100 has a first cylindrical section 100a extending upward from the mounting base 200. The first cylindrical section 100a is flared from bottom to top and is made of an elastic material so that when the cover plate closes the opening of each of the cylindrical bodies 100, the first cylindrical section 100a deforms and adheres to the cover plate. With this configuration, the first cylindrical section 100a essentially forms a suction cup structure, enabling it to adhere and fix itself to the cover plate when the cover plate closes the opening of the cylindrical body 100, thus strengthening the tight connection between the cover plate and the cylindrical body 100.

[0094] And / or, in one embodiment, the liquid forming cylinder 120 is provided with a filter screen 400 for filtering the liquid entering the liquid forming cylinder 120.

[0095] Furthermore, the liquid forming cylinder 120 has an inlet that communicates with other cylinders 100. When the inlet is set as one and positioned above the central axis of the liquid forming cylinder 120, the filter screen 400 can have a guide fluid protruding directly below the inlet. The guide fluid forms an annular guide surface, which gradually slopes upward or bends from its edge toward its center, so as to guide the liquid from the center of the annular guide surface to the periphery of the guide fluid, thereby fully dispersing the liquid before it flows into the inner cavity of the liquid forming cylinder 120.

[0096] When multiple feed inlets are provided, these multiple feed inlets can be arranged at intervals along the circumferential direction of the filter screen 400, which can also achieve the dispersion of liquid material into the inner cavity of the liquid forming cylinder 120.

[0097] Based on any of the above embodiments, the dissociation module 1 or the body may be provided with the cover plate. The cover plate at least covers each of the cylinders 100.

[0098] When the dissociation module 1 is equipped with the cover plate, the cover plate may be transparent at least at the opening of each of the cylinders 100, so that the dissociation situation inside each cylinder 100 can be visually observed by the experimenter. Of course, the cover plate may also be opaque, and the situation inside each cylinder 100 may be acquired and identified by, for example, an image recognition device.

[0099] When the cover plate is installed on the body, specifically, the body includes a base and a cover plate. The base is provided with an installation groove for the dissociation module 1 to be installed. The cover plate is movably installed on the cover plate so as to simultaneously open and close the groove of the installation groove and the opening of each of the cylinders 100.

[0100] The flip-up design of the cover plate allows air to enter from one side of the first cylindrical section 100a when, for example, each of the cylinders 100 is adsorbed and fixed on the cover plate by the first cylindrical section 100a, making it easier to separate the first cylindrical section 100a from the cover plate.

[0101] In a further embodiment, the portion of the cover plate used to cover each of the cylinders 100 may be provided with multiple sealing films. The multiple sealing films are stacked and arranged so that when the cover plate covers each of the cylinders 100, the sealing films are sealed to each cylinder 100. If the liquid forming cylinder 120 is sealed, it can be ensured that the obtained single-cell suspension is not contaminated by the external environment; if the remaining cylinders 100 are sealed, it can be ensured that the liquid remaining in the remaining cylinders 100 will not contaminate the external environment.

[0102] The sealing film can be a heat-pressed film or a self-adhesive film that meets the application scenario, and there are no restrictions.

[0103] The dissociation device can be based solely on the dissociation module 1 to achieve the purpose of dissociating tissue and obtaining a single-cell suspension; or, in addition to the dissociation module 1, the dissociation device may integrate other functional components for collecting, processing, or storing the single-cell suspension or various liquids required during the dissociation process.

[0104] Specifically, the dissociation device further includes a collection module, which is capable of collecting each single-cell suspension obtained by the dissociation module 1. For example, the single-cell suspensions can be classified and placed according to needs or types.

[0105] The dissociation device also includes a processing module, which can be configured to process the single-cell suspensions obtained by the dissociation module 1 according to actual needs. For example, it can be a centrifugation mechanism, a temperature control mechanism, etc.

[0106] The dissociation device also includes a storage module, which may be equipped with various sensors, such as temperature sensors, humidity sensors, and sterilization components, to provide a suitable storage environment for the single-cell suspensions obtained by the dissociation module 1.

[0107] The dissociation device also includes a feeding module or a waste collection module. The feeding module provides the dissociation module 1 with the required tissue, the required amount of lysis solution, lysis red solution and termination solution, etc.; the waste collection module can collect and process the waste generated after dissociation.

[0108] In one specific embodiment, the experimenter first prepares the tissue, for example, by adding PBS solution to a six-well plate to wash the sample stored in the tissue preservation solution; taking an appropriate amount of the washed sample and adding it to the dissociation solution, and cutting the tissue into tissue blocks of appropriate size using surgical scissors.

[0109] Next, the experimenter or the feeding module places the tissue into the dissociation cylinder 110, adjusts the temperature control mechanism to maintain the temperature inside the dissociation cylinder 110 at 37°C, and controls the cutter 300 to work for 15-40 minutes. Specifically, the cutter 300 needs to break up the tissue in the forward direction and then lift it in the reverse direction for the same amount of time every 5 minutes, ideally until the tissue is completely digested and forms a homogenized solution. Dissociation solution in the dissociation liquid container can be added to the dissociation cylinder 110 during this period.

[0110] Next, the homogenate obtained after lysis is filtered through a 40-70 μm sterile cell sieve. An equal volume of stop solution is added to the filtrate to terminate dissociation, yielding a cell suspension. In the dissociation chamber 110, or after transferring the cell suspension to the lysis chamber 120, an equal volume of erythrocyte lysis buffer is added. After inverting and mixing, the mixture is incubated at room temperature for 2-7 minutes. Then, an equal volume of stop solution is added to stop erythrocyte lysis. After inverting and mixing and removing the supernatant, a single-cell pellet is obtained, which is then resuspended to obtain a single-cell suspension.

[0111] The above description is merely a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural transformations made using the contents of the present invention's specification and drawings under the inventive concept of the present invention, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present invention.

Claims

1. A dissociation module for installation into a dissociation device, characterized in that, The dissociation module includes multiple cylinders for storing liquid and a mounting base connecting the multiple cylinders into one unit. The multiple cylinders include: The dissociation cylinder is equipped with a movably mounted cutter, which is used to break up the tissue inside the dissociation cylinder during its moving stroke; A liquid-forming cylinder, connected to the dissociation cylinder, is used to store the single-cell suspension obtained from dissociation; and... A storage tank for storing dissociation additives and connected to the dissociation tank and / or the liquid-forming tank; The dissociation module is connected to the power component in the dissociation device to drive the cutter movement and drive the liquid material to flow along the connection between the two cylinders via the power component. The two connected cylinders are a first cylinder and a second cylinder, respectively; The dissociation module further includes an installation cavity for the removable installation of the pressure regulating component in the power assembly. The installation cavity is connected to the first cylinder to adjust the pressure inside the first cylinder, thereby controlling the liquid to be drawn from the second cylinder into the first cylinder or discharged from the first cylinder into the second cylinder. The plurality of cylinders include a third cylinder, the third cylinder including a cylinder body and a bottom plate, the cylinder body having a cavity through it along its axial direction and forming two openings, the bottom plate being movably installed in the cavity along the axial direction of the cylinder body and being movably sealed with the inner wall of the cavity, so that under the drive of the telescopic component in the power assembly, the bottom plate is translated from one of the openings toward the other opening, and pushes out the liquid in the third cylinder; The cutting tool includes: The first cutter body includes a shaft rotatably disposed about the axis of the dissociation cylinder and a rib protruding from the side wall of the shaft. The shaft is connected to a rotary drive component in the power assembly, and the rib has a downward-facing first cutting face. The second blade body protrudes from the periphery of the shaft body below the rib, and the second blade body has a second blade surface facing upward. The second cutting surface is spaced apart from the first cutting surface, and the space defines a grinding area, so that during the rotation stroke of the first cutting body, the first cutting surface and the second cutting surface rotate relative to each other, and the tissue passing through the grinding area is ground. The upper end of the shaft is provided with a groove facing downwards, and a through hole is provided in the groove near the bottom wall of the dissociation cylinder. The groove is at least connected to the liquid forming cylinder.

2. The dissociation module as described in claim 1, characterized in that, The rib extends spirally along the circumferential and axial directions of the shaft. The helix angle of the rib gradually decreases from top to bottom; The height of the protruding ribs from the shaft body gradually decreases from bottom to top.

3. The dissociation module as described in claim 1, characterized in that, The second cutter body has multiple cutters arranged sequentially along the circumference of the shaft body; The second cutting face extends circumferentially and upward from the bottom wall of the dissociation cylinder towards the shaft body. The second cutting body also has a third cutting face connecting the top end of the second cutting face and the bottom wall of the dissociation cylinder. The inclination angle between the second cutting face and the bottom wall of the dissociation cylinder is not less than the inclination angle between the third cutting face and the bottom wall of the dissociation cylinder.

4. The dissociation module as described in claim 1, characterized in that, The cylinder has a first cylindrical section extending upward from the mounting base. The first cylindrical section is flared from bottom to top and is made of elastic material so that when the cover plate in the disintegration device covers the opening of each cylinder, the first cylindrical section deforms and adheres to the cover plate. The liquid forming cylinder is equipped with a filter screen, which is used to filter the liquid entering the liquid forming cylinder.

5. A dissociation device, characterized in that, include: main body; A power assembly is disposed on the main body; and, The dissociation module as described in any one of claims 1 to 4 is detachably connected to the main body and the power component, respectively.

6. The dissociation device as described in claim 5, characterized in that, The power assembly includes a pressure regulating component, a telescopic component, and a rotary drive component; The main body includes a base and a cover plate. The base is provided with an installation groove for installing the dissociation module. The cover plate is movably installed in the installation groove so as to simultaneously open and close the groove opening of the installation groove and the opening of each of the cylinders.

Images