Vessel for tissue treatment device and tissue treatment device

Abstract

The invention provides a vessel for a tissue treatment device, comprising: a sample chamber, the bottom of which forms a pyrolysis chamber; the whole cracking cavity is in a funnel shape, the cracking cavity is provided with a first liquid outlet, and the first liquid outlet is located in the bottom of the cracking cavity or located in the lower position of the side wall of the cracking cavity; the collecting cavity is lower than the cracking cavity; a second liquid inlet is formed in the top of the collecting cavity and is communicated with the first liquid outlet of the cracking cavity; the collecting cavity is provided with a second liquid outlet. The invention further provides a tissue treatment device, the tissue treatment device comprises the vessel for the tissue treatment device and the acoustic fluid device, the acoustic fluid device comprises an acoustic fluid chip, and the action range of the acoustic fluid chip in the working state comprises at least one part of the cracking cavity. According to the present invention, the deposition and the agglomeration of the cells in the lysis chamber can be reduced so as to improve the cell yield, and the method can further be used for the size screening collection, the negative selection, the positive selection, the cleaning, the enrichment, the extraction of the protein or the nucleic acid and the like of the cells.

Description

Technical Field

[0001] The present invention relates to the field of special sound waves and biotechnological processing, and particularly to a tissue processing device and a container for the device. Background Art

[0002] In biological scientific research experiments or in the medical clinical field, it is often necessary to rapidly lyse biological tissues into single-cell suspensions for further analysis of the single-cell suspensions. For example, a flow cytometer can be used to identify, count, and perform parameter analysis on the cells in the obtained single-cell suspension.

[0003] Among them, some traditional methods for preparing single-cell suspensions by lysis include: shaking table enzymatic digestion method and physical mechanical method. Among them, the digestion process of the shaking table enzymatic digestion method is relatively slow, about 1 to 2 hours, and the enzyme will affect cell activity. The physical mechanical methods such as cutting, sieving extrusion, pipetting with a syringe or pipette, etc., although fast, because their principle is to use cutting force to forcibly break the connections between cells in the tissue, so cells are extremely likely to die and break during this process, and the cell survival rate is low.

[0004] From the above, traditional lysis methods are either slow in lysis speed, low in cell activity, or cause large cell damage, and the effective cell yield (yield refers to the proportion of effective cells in all cells) is low. Based on this, a sono-fluidic device and an automated lysis and biopsy device for rapid tissue lysis are disclosed in the patent with Chinese Patent Publication No. CN115232724A. In this patent, a biological tissue sample is placed in a liquid, and a sono-fluidic effect is induced by the sono-fluidic device acting on the liquid, forming a forward jet and a vortex formed by the swirl acting on the biological tissue sample to achieve rapid lysis of the biological tissue into a single-cell suspension. This solution has a fast lysis speed, strong cell activity, small cell loss, and high cell yield.

[0005] And a further improvement based on the above solution is the technical problem to be solved by this application. Summary of the Invention

[0006] In view of this, the main purpose of the present invention is to provide a tissue processing device and a container to further improve the cell yield.

[0007] The first aspect of this application provides a vessel for a tissue processing device, including: a sample chamber, the bottom of the sample chamber forms a lysis chamber; the lysis chamber is funnel-shaped as a whole, the lysis chamber has a first liquid outlet, the first liquid outlet is located at the bottom of the lysis chamber, or the first liquid outlet is located at a lower position on the side wall of the lysis chamber; a collection chamber, arranged below the lysis chamber; the top of the collection chamber has a second liquid inlet, and the second liquid inlet is communicated with the first liquid outlet of the lysis chamber; the collection chamber has a second liquid outlet.

[0008] As described above, since the collection chamber is arranged lower than the sample chamber, and the first liquid outlet of the lysis chamber is located at the bottom of the lysis chamber or at a lower position on the side wall of the lysis chamber, when the solution processed by the lysis chamber is discharged downstream, the deposition of cells in the lysis chamber and the resulting agglomeration can be reduced, thereby further improving the cell yield. On the other hand, the shape of the lysis chamber acts on the acoustic fluidic effect of the acoustic fluidic chip described below, so that the vortices and jets generated by the acoustic fluidic effect can increase the overlapping range, improve the probability and effect that the particles in the vortices are simultaneously affected by the jets, and facilitate the lysis of tissue blocks in the solution.

[0009] As a possible implementation of the first aspect, the bottom of the collection chamber has a third liquid outlet; further includes a waste liquid chamber, which is arranged lower than the collection chamber; the top of the waste liquid chamber has a third liquid inlet, and the third liquid inlet is communicated with the third liquid outlet of the collection chamber; the waste liquid chamber has a fourth liquid outlet.

[0010] As described above, since the waste liquid chamber is arranged lower than the collection chamber, it is beneficial to the flow of the solution downstream (from the collection chamber to the waste liquid chamber).

[0011] As a possible implementation of the first aspect, it further includes a liquid collection chamber, the bottom of the liquid collection chamber forms the collection chamber; the liquid collection chamber is nested outside the sample chamber.

[0012] As described above, such a nested structure makes the overall structure more compact.

[0013] As a possible implementation of the first aspect, the second liquid outlet is located at a lower position on the side wall of the collection chamber.

[0014] As described above, the position of the second liquid outlet can effectively discharge the single-cell suspension in the collection chamber, reducing the possibility of deposition and agglomeration of single cells in the collection chamber.

[0015] As a possible implementation of the first aspect, a first filter membrane is arranged at the first liquid outlet of the lysis chamber, and a second filter membrane is arranged at the third liquid outlet of the collection chamber, and the pore diameter of the first filter membrane is larger than the pore diameter of the second filter membrane.

[0016] As described above, through the setting of the pore diameters of the first filter membrane and the second filter membrane, single cells of the desired size are retained in the collection chamber.

[0017] The second aspect of the present application provides a tissue processing device, including: any of the vessels for tissue processing devices provided in the first aspect of the present application; an acoustic fluidic device, including an acoustic fluidic chip, and the action range of the acoustic fluidic chip in the working state includes at least a part of the lysis chamber of the vessel of the tissue processing device.

[0018] As described above, tissue lysis is achieved through the acoustic fluidic chip, with fast lysis speed, strong cell viability, small cell loss, and high cell yield.

[0019] As a possible implementation of the second aspect, it further includes: a cover or support disposed at the orifice of the sample chamber, on which the main body of the acoustic fluidic device extending downward into the sample chamber is assembled, and the acoustic fluidic chip is located at the lower end of the main body of the acoustic fluidic device.

[0020] As a possible implementation of the second aspect, the main body of the acoustic fluidic device is disposed laterally facing the side wall of the lysis chamber; the acoustic fluidic chip is located at the end of the main body of the acoustic fluidic device facing the lysis chamber, and is in contact with or extends into the lysis chamber side wall.

[0021] As a possible implementation of the second aspect, the main body of the acoustic fluidic device is disposed horizontally facing the bottom of the lysis chamber; the acoustic fluidic chip is located at the bottom of the lysis chamber, and the acoustic fluidic chip does not block the first liquid outlet of the lysis chamber.

[0022] As described above, the setting method of the required acoustic fluidic device can be flexibly selected according to needs.

[0023] The third aspect of the present application provides a tissue processing method, which uses any of the above-mentioned utensils for tissue processing, or uses any of the above-mentioned devices for tissue processing.

[0024] As a possible implementation of the third aspect, the tissue processing includes at least one of the following:

[0025] Achieving tissue lysis in the lysis chamber;

[0026] Cooperating with the filtration of the first filter membrane and the second filter membrane to achieve cell size screening, and collecting target-sized cells from the collection chamber;

[0027] Using antibody-modified magnetic beads or microspheres as carriers to bind target cells to the carriers, and cooperating with the filtration of the first filter membrane and the second filter membrane to perform negative or positive selection of target cells;

[0028] Cooperating with the filtration of the first filter membrane and the second filter membrane to perform solution replacement to achieve washing of the cells in the collection chamber;

[0029] Cooperating with the filtration of the first filter membrane and the second filter membrane to achieve enrichment of target cells;

[0030] Cooperating with microspheres as carriers to bind proteins or nucleic acids to the carriers, and cooperating with the filtration of the first filter membrane and the second filter membrane to perform extraction of proteins or nucleic acids. Description of the Drawings

[0031] Figure 1 Schematic structural diagram of a tissue processing device provided for the third embodiment of the present application;

[0032] Figure 2 Schematic structural diagram of a tissue processing device provided for the fourth embodiment of the present application;

[0033] Figure 3 Schematic structural diagram of a tissue processing device provided for the fifth embodiment of the present application;

[0034] Figure 4 For the present application Figure 2 Schematic diagram showing a usage state of the tissue processing device shown;

[0035] Figure 5 Schematic diagram of a device including a tissue processing device for the present application;

[0036] Figure 6 Another schematic diagram of a device including a tissue processing device for the present application.

[0037] Description of reference numerals:

[0038] Sample chamber 10, lysis chamber 11, first liquid outlet 12, collection chamber 20, second liquid inlet 21, second liquid outlet 22, third liquid outlet 23, liquid collection chamber 24, fifth liquid outlet 25, waste liquid chamber 30, third liquid inlet 31, fourth liquid outlet 32, interface 33 of external pipeline, acoustic fluid device 40, plug 50. Detailed implementation manners

[0039] The following are embodiments given in conjunction with the accompanying drawings to further illustrate the technical solutions provided by the present application. It should be understood that the system structure and service scenarios provided in the embodiments of the present application are mainly for illustrating possible implementation manners of the technical solutions of the present application, and should not be construed as the only limitation to the technical solutions of the present application. Those of ordinary skill in the art can know that with the evolution of the system structure and the emergence of new service scenarios, the technical solutions provided by the present application are equally applicable to similar technical problems.

[0040] It should be understood that the tissue processing solutions provided in the embodiments of the present application include a tissue processing device, a container for the tissue processing device, and a detection device including the tissue processing device. Since the principles of these technical solutions for solving problems are the same or similar, in the following introduction of specific embodiments, some repetitions may not be elaborated again, but it should be regarded that there are mutual references between these specific embodiments and they can be combined with each other.

[0041] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this application belongs. In case of inconsistency, the meaning stated in this specification or the meaning derived from the content recorded in this specification shall prevail. Additionally, the terms used herein are only for the purpose of describing the embodiments of this application and are not intended to limit this application.

[0042] In the technology disclosed in Chinese Patent Publication No. CN115232724A, an automated lysis and biopsy device is disclosed. Refer to the schematic diagram of the automated lysis and biopsy device for rapid tissue lysis disclosed in this patent, or refer to the three-dimensional diagram of the automated lysis and biopsy device for rapid tissue lysis disclosed in this patent. The bottom of the liquid inlet pipe, sample inlet pipe, sample outlet pipe, and waste liquid pipe included in the device are connected by a horizontal pipeline. A lysis chamber lower than the horizontal pipeline is provided at the bottom of the sample inlet pipe to serve as the working area of the acoustofluidic device. Among them, when the acoustofluidic device is turned off after the lysis process is completed, during the process of injecting culture medium from the injection pipe and sucking out the solution from the waste liquid pipe, since the culture medium flows horizontally along the bottom horizontal pipeline, some cells (including single cells) deposited in the lysis chamber below the horizontal pipeline are difficult to move downstream along the fluid direction, and the deposited cells may further agglomerate, which will affect the cell yield.

[0043] The improved solution provided by this application can effectively reduce cell deposition and the resulting agglomeration, thereby further improving the cell yield.

[0044] The solution provided by this application is used for tissue lysis, and the result after lysis is a single cell suspension, which can be used for subsequent cell manipulation or analysis, such as cell staining, cell biopsy, cancer cell screening, single cell manipulation, extraction or assembly, etc.

[0045] The technical solution of this application will be described below.

[0046] The first embodiment of this application provides a vessel for a tissue processing device, including:

[0047] A sample chamber 10, the bottom of the sample chamber 10 constitutes a lysis chamber 11;

[0048] The orifice at the top of the sample chamber 10 can be an interface 33 for an external pipeline. In some embodiments, the external pipeline can be installed on a cover body, and the cover body can be assembled at the interface 33 of the external pipeline. The cover body can be in the shape of a plug, a cap, etc. The cover body can be made of a material with certain elasticity, which is convenient for the insertion, extraction, and replacement of one or more external pipelines. In some embodiments, the cover body is also used to fix the main body of the acoustofluidic device 40 extending into the sample chamber (see the description later, not elaborated here).

[0049] The cracking chamber 11 is generally funnel-shaped as a whole. The cracking chamber 11 has a first liquid outlet 12, and the first liquid outlet 12 is located at the bottom of the cracking chamber 11, or the first liquid outlet 12 is located at a lower position on the side wall of the cracking chamber 11.

[0050] A collection chamber 20 is provided below the cracking chamber 11. The top of the collection chamber 20 has a second liquid inlet 21, and the second liquid inlet 21 is communicated with the first liquid outlet 12 of the cracking chamber 11. The collection chamber 20 has a second liquid outlet 22.

[0051] From the above, since the collection chamber 20 is provided below the sample chamber 10, and the first liquid outlet 12 of the cracking chamber 11 is located at the bottom of the cracking chamber 11 or at a lower position on the side wall of the cracking chamber 11, when the solution processed by the cracking chamber 11 is discharged downstream, the deposition of cells in the cracking chamber 11 and the resulting agglomeration can be reduced, thereby further improving the cell yield.

[0052] In some embodiments, the sample chamber 10 can be cylindrical, such as a circular cylinder or a polygonal ring cylinder (referring to a cylinder whose cross-section perpendicular to the cylinder axis is a polygon). The polygonal ring cylinder is, for example, a quadrilateral ring cylinder or a hexagonal ring cylinder. In some embodiments, the sample chamber 10 can also be an irregular cylinder, such as a cylinder with a gradually decreasing inner diameter from top to bottom.

[0053] In some embodiments, the cracking chamber 11 is generally funnel-shaped as a whole, and can be a cylinder with a gradually decreasing inner peripheral circumference from top to bottom. For example, the cracking chamber 11 forms a cavity similar to a frustum of a cone, a frustum of a pyramid or other frustums of a pyramid, a pear shape, etc. In some embodiments, when the bottom of the sample chamber 10 forms the cracking chamber 11, it can be smoothly formed into the cracking chamber 11. Among them, through the shape of the cracking chamber 11 acting on the acoustic fluid effect of the subsequent acoustic fluid chip, the vortex and jet generated by the acoustic fluid effect can increase the overlapping range, improve the probability and effect of the particles in the vortex being simultaneously affected by the jet, and are more conducive to the cracking of tissue blocks in the solution.

[0054] In some embodiments, the bottom of the collection chamber 20 has a third liquid outlet 23. The vessel further includes a waste liquid chamber 30 provided below the collection chamber 20. The top of the waste liquid chamber 30 has a third liquid inlet 31, and the third liquid inlet 31 is communicated with the third liquid outlet 23 of the collection chamber 20. The waste liquid chamber 30 has a fourth liquid outlet 32.

[0055] In some embodiments, the collection chamber 20 or the waste liquid chamber 30 may be cylindrical, such as a circular cylinder or a multi-sided annular cylinder (referring to a cylinder whose cross-section perpendicular to the cylinder axis is a polygon). The multi-sided annular cylinder may be, for example, a four-sided annular cylinder, a six-sided annular cylinder, etc. In some embodiments, the collection chamber 20 or the waste liquid chamber 30 may also be an irregular cylinder, such as a cylinder whose inner diameter gradually decreases from top to bottom.

[0056] In some embodiments, when the first liquid outlet 12 is provided at the bottom of the cracking chamber 11, the first liquid outlet 12 may be disposed directly opposite to the second liquid inlet 21 at the top of the collection chamber 20. In some embodiments, when the third liquid outlet 23 is provided at the bottom of the collection chamber 20, the third liquid outlet 23 may be disposed directly opposite to the third liquid inlet 31 at the top of the waste liquid chamber 30.

[0057] In some embodiments, a liquid collection chamber 24 is further included, and the bottom of the liquid collection chamber 24 forms the collection chamber 20; the liquid collection chamber 24 is nested outside the sample chamber 10. In some embodiments, the sample chamber 10 can be detachably placed into the liquid collection chamber 24 or taken out from the top opening of the liquid collection chamber 24.

[0058] In some embodiments, an image acquisition device may also be provided, which is disposed facing the collection chamber 20. In some embodiments, the image acquisition device may be disposed below the collection chamber 20 to acquire an image of the upper collection chamber 20.

[0059] In some embodiments, the image acquisition device and the waste liquid chamber 30 may be disposed together below the collection chamber 20. In some embodiments, the third liquid outlet 23 at the bottom of the collection chamber 20 is located near the side wall at its bottom, and the waste liquid chamber 30 is disposed deviating from the bottom of the collection chamber 20. For example, the waste liquid chamber 30 is tangent to the collection chamber 20 in the vertical downward projection, and the image acquisition device is disposed directly opposite to the center of the bottom of the collection chamber 20, so that the image acquisition device can acquire a complete image of the collection chamber 20 as much as possible.

[0060] In some embodiments, the image acquisition device may include a camera or an image sensor, etc. In some embodiments, the camera or the image sensor may be the camera or the image sensor of an electronic device, and the electronic device may be, for example, an electron microscope, a mobile phone, a photographic equipment, etc.

[0061] In some embodiments, the second liquid outlet 22 of the collection chamber 20 is disposed at a lower position on the side wall of the collection chamber 20 to facilitate the discharge of the liquid in the collection chamber 20 from the second liquid outlet 22.

[0062] In some embodiments, a first filter membrane is provided at the first liquid outlet 12 of the cracking chamber 11, and a second filter membrane is provided at the third liquid outlet 23 of the collection chamber 20. The pore size of the first filter membrane is larger than that of the second filter membrane, so as to facilitate the collection of particulate matter of a desired size in the collection chamber 20. In some embodiments, the pore size of the first filter membrane is 70-100 μm, and the pore size of the second filter membrane is 30-50 μm.

[0063] In some embodiments, the above-mentioned sample chamber 10, collection chamber 20 and waste liquid chamber 30 may be made of resin or other materials.

[0064] The second embodiment of the present application provides a tissue processing device, including the solution of the vessel for the tissue processing device described in the first embodiment or the solution described in any optional embodiment thereof, and an acoustic fluid device 40. The acoustic fluid device 40 includes an acoustic fluid chip, and the working range of the acoustic fluid chip includes at least a part of the cracking chamber 11.

[0065] In some embodiments, a cover or a support is further provided at the opening of the sample chamber 10. The main body of the acoustic fluid device 40 extending downward into the sample chamber 10 is assembled on the cover or the support, and the acoustic fluid chip is located at the lower end of the main body of the acoustic fluid device 40.

[0066] In some embodiments, the cover can close or semi-close the opening of the sample chamber 10. The main body of the acoustic fluid device 40 is inserted and assembled through a through hole on the cover, and the insertion depth or the angle of fine adjustment offset from the vertical direction can be adjusted to adjust the position of the acoustic fluid chip in the cracking chamber 11.

[0067] In some embodiments, the support can be a bracket, and in other embodiments, the support can be a displacement stage, such as a displacement stage with a horizontal movement range or a three-dimensional displacement stage with a spatial movement range.

[0068] In some embodiments, the opening of the sample chamber can also be used as an interface 33 for an external pipeline. In some embodiments, the external pipeline can be installed on a cover, and the cover can be assembled at the interface 33 of the external pipeline. The cover can be in the shape of a plug, a cap, etc. The cover can be made of a material with a certain elasticity, which is convenient for the insertion, extraction and replacement of one or more external pipelines, and is also convenient for fixing and sealing the interface 33 of the external pipeline. In some embodiments, the cover is used to fix the main body of the acoustic fluid device 40 extending into the sample chamber. In some embodiments, one or more external pipelines can be inserted or replaced on the cover, or it is only used for sealing function.

[0069] Among them, according to the functions to be achieved by the tissue processing device, multiple external pipelines may need to be inserted simultaneously at the interface 33 of an external pipeline, or one or more external pipelines may need to be inserted simultaneously at the interfaces 33 of external pipelines at different positions. Among them, the external pipeline can be any of the following:

[0070] 1) It can be a pipeline communicating with the atmosphere. The outer end of the pipeline can be opened (opened means communicating with the atmosphere) or closed as needed. For example, a valve is provided at the outer end of the pipeline, or a detachable plug is used, or a clip is used to fold the outer end of the pipeline in half and clamp it closed;

[0071] 2) It can be a pipeline communicating with a liquid supply device. The liquid supply device can provide an enzyme solution, or a target solution to be processed, or a replacement solution, etc.;

[0072] 3) It can be a pipeline communicating with an air pump, so as to introduce gas or pump air, so as to form a positive pressure or a negative pressure in the tissue processing device, so as to affect the flow of the liquid in the tissue processing device;

[0073] 4) It can be a detachable plug for closing, which cooperates with the pipelines of the air pumps at other positions;

[0074] 5) It can be a pipeline communicating with a pump to pump out the liquid, usually assembled at the interface 33 of the external pipeline on the side of the waste liquid chamber 30.

[0075] In some embodiments, the main body side of the acoustic fluid device 40 faces the side wall of the lysis chamber 11; the acoustic fluid chip is located at the end of the acoustic fluid device 40, abuts against the side wall of the lysis chamber 11 or extends into the lysis chamber 11.

[0076] In some embodiments, when the main body side of the acoustic fluid device 40 faces the side wall of the lysis chamber 11, it can be horizontal or approximately horizontal, or the main body of the acoustic fluid device 40 is arranged at an angle to the horizontal.

[0077] In some embodiments, the main body of the acoustic fluid device 40 faces the bottom of the lysis chamber 11 horizontally; the acoustic fluid chip is located at the bottom of the lysis chamber 11, and the acoustic fluid chip does not block the first liquid outlet 12, and the working surface of the acoustic fluid chip faces the upper lysis chamber 11.

[0078] In some embodiments, the structure in which the acoustic fluid chip does not block the first liquid outlet 12, for example: the acoustic fluid chip and the first liquid outlet 12 are arranged in parallel at the bottom of the lysis chamber 11; another example: at the bottom of the lysis chamber 11, the first liquid outlet 12 partially surrounds or encircles the acoustic fluid chip; still another example: the first liquid outlet 12 is a sieve-like structure including a plurality of through holes, and the acoustic fluid chip is located on the sieve-like structure and does not block all the through holes on the sieve-like structure.

[0079] In some embodiments, the main body of the acoustic-fluidic device 40 may be rod-shaped, and the acoustic-fluidic chip is located at its end. In other embodiments, the main body of the acoustic-fluidic chip may be linear or sheet-shaped, such as a flexible sheet formed by a flexible cable.

[0080] In some embodiments, when the main body of the acoustic-fluidic device 40 is rod-shaped, it may include a housing, a PCB board located at one end of the housing, and a terminal at the other end. The PCB board is electrically connected to the terminal through an RF connecting wire. The acoustic-fluidic chip is disposed on the side of the PCB board facing outward and is circuit-connected to the PCB. The PCB mainly functions to transfer signal lines and ground lines.

[0081] In some embodiments, the acoustic-fluidic chip may be single or multiple. In some embodiments, the acoustic-fluidic chip includes a surface acoustic wave resonator chip, a bulk acoustic wave resonator chip, or a piezoelectric ceramic chip. Among them, when it is a bulk acoustic wave resonator chip, it can be a solidly mounted bulk acoustic wave resonator chip; the resonance frequency range of the solidly mounted bulk acoustic wave resonator is 1 GHz to 10 GHz, and the applied power range is 0.1 W to 10 W.

[0082] In some embodiments, there may be multiple acoustic-fluidic devices 40. In some embodiments, when there are multiple ones, they can be arranged in any of the following single or combined manners: the main bodies of some acoustic-fluidic devices 40 may be laterally oriented towards the side wall of the cracking chamber 11, the main bodies of some acoustic-fluidic devices 40 may be horizontally oriented towards the bottom of the cracking chamber 11, some acoustic-fluidic devices 40 may be vertically inserted into the cracking chamber 11, and so on.

[0083] Next, with further reference to the drawings, some specific embodiments of the present application will be introduced to better understand the technical solution of the present application. Among them, the following specific embodiments can be adapted to different scenario requirements respectively.

[0084] Such as Figure 1Fig. 0 shows a tissue processing device provided by the third embodiment of the present application. The tissue processing device of this embodiment includes a sample chamber 10 containing a lysis chamber 11, a collection chamber 20, and a waste liquid chamber 30, which are arranged in sequence from top to bottom. The main body of the acoustic fluid device 40 is horizontally oriented towards the lysis chamber 11, and the acoustic fluid chip at the end of the main body of the acoustic fluid device 40 is attached to the side wall of the lysis chamber 11. The orientation of the second liquid outlet 22 of the collection chamber 20 is perpendicular to the extending direction of the main body of the acoustic fluid device 40 in space. The fourth liquid outlet 32 of the waste liquid chamber 30 extends upward from the top of the waste liquid chamber 30 and is arranged side by side with the sample chamber 10, and the top of the sample chamber 10 is arranged side by side with the top of the fourth liquid outlet 32 of the waste liquid chamber 30. Among them, the top opening of the sample chamber 10 can be used as an interface 33 for an external pipeline, and the top opening of the fourth liquid outlet 32 of the waste liquid chamber 30 can be used as an interface 33 for an external pipeline to connect or replace different external pipelines as needed. Among them, there is a first filter membrane between the first liquid outlet 12 of the lysis chamber 11 and the second liquid inlet 21 of the collection chamber 20, and there is a second filter membrane between the third liquid outlet 23 of the collection chamber 20 and the third liquid inlet 31 of the waste liquid chamber 30.

[0085] Among them, when using Figure 1 the embodiment shown for tissue processing, most of the over-sized particles will be retained in the lysis chamber 11 before being filtered by the first filter membrane, and most of the undersized particles will flow through the second filter membrane to the waste liquid chamber 30, so that a single cell suspension retained in the collection chamber 20 after being filtered by the first filter membrane and before the second filter membrane can be obtained, and the single cells therein are mainly single cells of the desired size.

[0086] As Figure 2 Fig. 10 shows a tissue processing device provided by the fourth embodiment of the present application, which includes a sample chamber 10 containing a lysis chamber 11, a collection chamber 20, and a waste liquid chamber 30, which are arranged in sequence from top to bottom. The side wall of the lysis chamber 11 is connected to the top of the collection chamber below through a channel, which is a bent channel in the figure. Based on the manufacturing process, a plug 50 is shown in the figure to form the bent channel. The second liquid outlet 22 of the collection chamber 20 is horizontally oriented. The fourth liquid outlet 32 of the waste liquid chamber 30 is arranged near the bottom of the side wall of the waste liquid chamber 30, and the waste liquid chamber 30 also has another opening at its top for use as an interface 33 for an external pipeline. Among them, there is a first filter membrane between the first liquid outlet 12 of the lysis chamber 11 and the second liquid inlet 21 of the collection chamber 20, and there is a second filter membrane between the third liquid outlet 23 of the collection chamber 20 and the third liquid inlet 31 of the waste liquid chamber 30. In this embodiment, the acoustic fluid chip of the acoustic fluid device 40 can be located at the bottom of the lysis chamber 11 or can be arranged on the cover body and extend into the lysis chamber 11 from the upper end opening of the sample chamber 10. Among them, the top opening of the sample chamber 10 can be used as an interface 33 for an external pipeline, and the other opening connecting the top of the waste liquid chamber 30 is used as an interface 33 for an external pipeline to connect or replace different external pipelines as needed.

[0087] As Figure 4 shown, a picture of a usage state of a device based on Figure 2 is presented. Figure 4 In Figure 4 , a cover is assembled on the interface 33 of the external pipeline at the top of the sample chamber 10. A hose communicating with the atmosphere (the end of which can be opened or closed as needed) is inserted into the cover, and a hose communicating with the liquid supply device is also inserted. A hose communicating with an air pump is inserted into the cover on the interface 33 of the external pipeline at the top of the sample chamber 10. Only the control process of liquid flow is described below: In this example, the second liquid outlet 22 of the collection chamber 20 is temporarily closed, and the fourth liquid outlet 32 of the waste liquid chamber 30 is temporarily closed. Among them, when the hose communicating with the atmosphere is open and the air pump is cut off, the liquid supply device can supply liquid into the lysis chamber of the sample chamber 10 through the hose. When the hose communicating with the atmosphere is open and the air pump is pumping air, the liquid in the lysis chamber, or the liquid supplied by the liquid supply device simultaneously, can move downstream to the collection chamber 20 and the waste liquid chamber 30. When the hose communicating with the atmosphere is closed, the liquid supply device is cut off, and at this time the fourth liquid outlet 32 of the waste liquid chamber 30 is opened, when the air pump injects gas, the liquid in the waste liquid chamber 30 can be discharged from the fourth liquid outlet 32.

[0088] Among them, after tissue processing using Figure 2 the shown embodiment, most of the too large particles will be retained in the lysis chamber 11 before the first filter membrane filtration, and most of the too small particles will flow through the second filter membrane to the waste liquid chamber 30, so that a single cell suspension retained in the collection chamber 20 after the first filter membrane filtration and before the second filter membrane can be obtained, and the single cells therein are mainly single cells of the desired size.

[0089] As Figure 3 shown, a tissue processing device provided by the fifth embodiment of the present application is presented, including a sample chamber 10 containing a lysis chamber 11, a collection chamber 20, and a liquid collection chamber 24, which are sequentially arranged from top to bottom. The bottom of the liquid collection chamber 24 constitutes the collection chamber 20. The liquid collection chamber 24 is nested outside the sample chamber 10. The liquid collection chamber 24 has a fifth liquid outlet 25 communicating with the liquid collection chamber 24 (here the fifth liquid outlet 25 is equivalent to the second liquid outlet 22 of the aforementioned collection chamber 20), and the position of the fifth liquid outlet 25 is higher than the position of the main body of the acoustic fluid device 40. The main body of the acoustic fluid device 40 horizontally passes through the liquid collection chamber 24 and is arranged towards the lysis chamber 11. The acoustic fluid chip at the end of the main body of the acoustic fluid device 40 is attached to the side wall of the lysis chamber 11. Among them, there is a first filter membrane between the first liquid outlet 12 of the lysis chamber 11 and the second liquid inlet 21 of the collection chamber 20. Among them, the top opening of the sample chamber 10 can serve as the interface 33 of the external pipeline. In some cases, the fifth liquid outlet 25 of the liquid collection chamber 24 can also serve as the interface 33 of the external pipeline to connect or replace different external pipelines as needed.

[0090] Among them, after using Figure 3 the illustrated embodiment for tissue processing, a suspension containing single cells after filtration by the first filter membrane can be obtained. The suspension contains single cells of a desired size and may also include some particles not larger than single cells, such as cell debris, etc.

[0091] Next, the working principle of the tissue processing device of the present application will be described. Among them, taking the tissue processing device provided by the third embodiment of the present application to process tissue as an example, when performing tissue processing, the following steps are included:

[0092] S10: Place the tissue block to be lysed into the sample chamber 10, and inject an enzyme solution into the sample chamber 10 so that the tissue block to be lysed is immersed in the enzyme solution and deposited in the lysis chamber 11.

[0093] S20: If the acoustic fluidic device 40 is located on the cover or support, cover the opening of the sample chamber 10 with the cover or support so that the acoustic fluidic chip at the end of the acoustic fluidic device 40 extends downward into the lysis chamber 11 and is immersed in the enzyme solution. If the acoustic fluidic device 40 has been pre-set to be horizontally oriented towards the lysis chamber 11, this step can be ignored.

[0094] Among them, the distance between the acoustic fluidic chip and the tissue block in the lysis chamber 11 can be within 1 cm.

[0095] S30: Turn on the acoustic fluidic device 40. The acoustic fluidic chip acts on the lysis chamber 11. Under the combined action of the shape of the lysis chamber 11 and the bulk acoustic wave effect generated by the acoustic fluidic chip, the solution in the lysis chamber 11 generates eddy currents and jets, which act on the tissue block. The tissue block is captured in the vortex of the eddy current and rotates and moves under the combined action of the eddy current and the jet, and is impacted by the jet force. Through rotation, movement, and impact, the enzymolysis products are quickly stripped from the surface of the tissue block, exposing deeper stroma, thereby further accelerating the enzymolysis speed and achieving layer-by-layer stroma digestion and cell stripping. In this example, the acoustic fluidic device 40 can be kept working for a period of time, such as 10 - 20 min.

[0096] S40: After controlling the acoustic fluidic device 40 to stop working, inject cell culture medium into the sample chamber 10 (the above-mentioned cover or support covering the opening of the sample chamber 10 can be opened for injecting the cell culture medium, or a pipeline connected to the culture medium supply can be inserted into the cover on the top of the sample chamber 10 for injecting the cell culture medium), and at the same time suck out the solution from the waste liquid chamber 30 (the pump connected to the pipeline on the cover of the interface 33 of the external pipeline assembled on the side of the waste liquid chamber 30 can be turned on to pump out the liquid), for a period of time, such as 2 - 5 min, so that the enzyme solution is replaced by the cell culture medium.

[0097] In this process, the solution will sequentially pass through the first filter membrane at the first liquid outlet 12 of the lysis chamber 11 and the second filter membrane at the third liquid outlet 23 of the collection chamber 20 along the fluid direction. Particles larger than the desired size are intercepted by the first filter membrane in the lysis chamber 11, and particles smaller than the desired size flow through the second filter membrane towards the waste liquid chamber 30. Particles of the desired size are intercepted by the second filter membrane in the collection chamber 20. In this example, the particles of the desired size are single cells of the corresponding size, and these single cells are intercepted in the collection chamber 20. At this time, the collection chamber 20 contains a single cell suspension.

[0098] S50: Open the third liquid outlet 23 of the collection chamber 20 to obtain the single cell suspension flowing out of the collection chamber 20.

[0099] Among them, the outlet of the waste liquid chamber 30 can be closed (the pump connected to the pipeline on the cover of the interface 33 of the external pipeline assembled on the side of the waste liquid chamber 30 can be turned off, or the valve of this pipeline can be closed), the third liquid outlet 23 of the collection chamber 20 is opened, a closed structure is formed in the waste liquid chamber 30, and the cell culture solution or replacement solution is continuously injected into the sample chamber 10. The liquid flowing into the collection chamber 20 will carry the particles of the desired size therein and flow out along the third liquid outlet 23.

[0100] Among them, in the above step S10, injecting the enzyme solution into the sample chamber 10 is an optional step. In some other embodiments, the cell culture solution can also be injected. Injecting the enzyme solution can accelerate the lysis process through the treatment of the enzyme.

[0101] Among them, the present application also provides some embodiments, which are methods for tissue treatment, using any of the above-mentioned utensils for tissue treatment, or using any of the above-mentioned devices for tissue treatment. Specifically, the tissue treatment includes at least one of the following:

[0102] Realize the lysis of the tissue in the lysis chamber;

[0103] Cooperate with the filtration of the first filter membrane and the second filter membrane to realize the size screening of cells, and collect the target size cells from the collection chamber;

[0104] Use antibody-modified magnetic beads or microspheres as carriers to bind the target cells to the carriers, and cooperate with the filtration of the first filter membrane and the second filter membrane to perform negative selection or positive selection of the target cells;

[0105] Cooperate with the filtration of the first filter membrane and the second filter membrane to perform the replacement of the solution, and realize the cleaning of the cells in the collection chamber;

[0106] Cooperate with the filtration of the first filter membrane and the second filter membrane to realize the enrichment of the target cells;

[0107] With microspheres as carriers, proteins or nucleic acids are bound to the carriers, and protein or nucleic acid extraction is carried out in conjunction with the filtration of the first filter membrane and the second filter membrane.

[0108] Figure 5 This is a schematic diagram of a device including a tissue processing device according to the present application; Figure 6 This is another schematic diagram of a device including a tissue processing device according to the present application. Among them, it is a schematic diagram of hiding the outer casing, and the figure is a product schematic diagram.

[0109] The terms "first, second, third, etc." or similar terms such as module A, module B, module C, etc. in the description and claims are only used to distinguish similar objects and do not represent a specific order for the objects. Understandably, the specific order or sequence can be interchanged when permitted, so that the embodiments of the present application described here can be implemented in an order other than that illustrated or described here.

[0110] In the above description, the reference numerals representing steps, such as S110, S120... etc., do not necessarily mean that the steps will be executed in this order. The order of the front and rear steps can be interchanged when permitted, or they can be executed simultaneously.

[0111] The term "comprising" used in the description and claims should not be construed as being limited to the content listed thereafter; it does not exclude other elements or steps. Therefore, it should be construed as specifying the presence of the recited features, integers, steps or components, but not excluding the presence or addition of one or more other features, integers, steps or components and their groups. Therefore, the expression "a device comprising device A and B" should not be limited to a device consisting only of components A and B.

[0112] The "one embodiment" or "embodiment" mentioned in this specification means that the specific features, structures or characteristics described in connection with the embodiment are included in at least one embodiment of the present application. Therefore, the phrases "in one embodiment" or "in an embodiment" that appear throughout this specification do not necessarily all refer to the same embodiment, but may refer to the same embodiment. In addition, in one or more embodiments, the various specific features, structures or characteristics can be combined in any suitable manner, as will be apparent to those of ordinary skill in the art from this disclosure.

[0113] Note that the above is only a preferred embodiment of the present application and the technical principles applied. Those skilled in the art will understand that the present application is not limited to the specific embodiments described herein, and various obvious changes, re-adjustments and substitutions can be made by those skilled in the art without departing from the protection scope of the present application. Therefore, although the present application has been described in more detail through the above embodiments, the present application is not limited to the above embodiments only. Without departing from the concept of the present application, more other equivalent embodiments can be included, all of which fall within the protection scope of the present application.

Claims

1. A vessel for a tissue processing device, characterized in that: include: A sample chamber, wherein the bottom of the sample chamber forms a lysis chamber; The lysis chamber is funnel-shaped as a whole, and has a first liquid outlet, which is located at the bottom of the lysis chamber, or at a lower position of a side wall of the lysis chamber; The collecting chamber is arranged below the lysing chamber; the top of the collecting chamber is provided with a second liquid inlet, the second liquid inlet is communicated with the first liquid outlet of the lysing chamber; the collecting chamber has a second liquid outlet.

2. The vessel according to claim 1, characterized in that Also includes: The bottom of the collecting chamber is provided with a third liquid outlet; The waste liquid chamber is arranged below the collecting chamber; the top of the waste liquid chamber is provided with a third liquid inlet, the third liquid inlet is connected with the third liquid outlet of the collecting chamber; the waste liquid chamber is provided with a fourth liquid outlet.

3. The vessel according to claim 1, characterized in that Also includes: A liquid collecting chamber, the bottom of which constitutes the collecting chamber; The liquid collection chamber is nested outside the sample chamber.

4. The vessel according to claim 1, characterized in that The second liquid outlet is located at a lower position of the side wall of the collecting chamber.

5. The vessel according to claim 1, characterized in that A first filter membrane is arranged at the first liquid outlet of the lysis chamber, and a second filter membrane is arranged at the third liquid outlet of the collection chamber. The filter pore diameter of the first filter membrane is larger than the filter pore diameter of the second filter membrane.

6. A tissue processing device, characterized in that: include: The vessel for a tissue processing device according to any one of claims 1 to 5; The acoustic fluid device comprises an acoustic fluid chip, wherein the action range of the acoustic fluid chip in a working state includes at least a part of the lysis chamber of the vessel of the tissue processing device.

7. The device according to claim 6, characterized in that Also includes: A cover or support is arranged at the opening of the sample cavity, and the cover or support is equipped with a body of the acoustic fluid device extending downward into the sample cavity, and the acoustic fluid chip is located at the lower end of the body of the acoustic fluid device.

8. The device according to claim 6, characterized in that: The main body of the acoustic fluidic device is arranged laterally toward the side wall of the lysis chamber; The acoustic fluid chip is located at the end of the main body of the acoustic fluid device facing the lysis chamber, and is arranged in contact with the side wall of the lysis chamber or extends into the lysis chamber.

9. The device according to claim 6, characterized in that: The main body of the acoustic fluidic device is horizontally arranged toward the bottom of the lysis chamber; The acoustic fluid chip is located at the bottom of the lysis chamber, and the acoustic fluid chip does not close the first liquid outlet of the lysis chamber.

10. A tissue processing method, characterized in that: Use the vessel described in any one of claims 1 to 5 to perform tissue processing, or use the device described in any one of claims 6 to 9 to perform tissue processing.

11. The method of claim 10, wherein the tissue processing comprises at least one of the following: Achieving tissue lysis in a lysis chamber; Cooperating with the first filter membrane and the second filter membrane to filter, the cells are sized and collected from the collection chamber. Using antibody-modified magnetic beads or microspheres as carriers, binding target cells to the carriers, and performing negative or positive selection of target cells in cooperation with the filtration of the first filter membrane and the second filter membrane; Cooperating with the filtering of the first filter membrane and the second filter membrane, the solution is replaced to achieve the cleaning of the cells in the collection chamber; Coordinating the filtration of the first filter membrane and the second filter membrane to achieve enrichment of target cells; The microspheres are used as carriers to bind proteins or nucleic acids to the carriers, and the first and second filter membranes are used for filtration to extract proteins or nucleic acids.

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