Apparatus and method for processing biological sample

Abstract

The present application relates to a cartridge for processing a biological sample, such as a tissue or tumor sample. The cartridge may interface with a system, such as an automated system, to process a biological sample. The cartridges and / or systems of the present disclosure may be used in methods of processing biological samples. In certain use cases, the cartridges and / or systems and / or methods of the present disclosure may shred a biological sample. In certain use cases, the cartridges and / or systems and / or methods of the present disclosure may dissociate biological samples. In certain use cases, the cartridges and / or systems and / or methods of the present disclosure may homogenize biological samples.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 387,891, filed on December 16, 2022, the content of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to devices and methods for processing biological samples such as tissue samples. More specifically, the present disclosure relates to automated devices and methods for processing biological samples such as tissue samples. Even more specifically, the present disclosure relates to automated devices and methods for dissociating and / or homogenizing biological samples such as tissue samples. Background Art

[0004] In various cell and molecular biology applications, researchers and technicians typically need to process biological samples before performing downstream assays or analyses. In the case of cell biologists, cells or cell organelles obtained from solid tissues or tumors are typically used for assays. In the case of molecular biologists, nucleic acids and proteins obtained from cells such as those located in liquid samples or solid tissue or tumor samples are typically used for assays.

[0005] It is common to obtain cell suspensions from tissues or tumors using enzymes or enzyme - containing solutions. In fact, many commercial suppliers provide reagents for this purpose. However, depending on the tissue or tumor type, enzymatic dissociation may require a long period of time and / or result in incomplete dissociation and / or digestion of related substances such as proteins or glycoproteins. Less commonly, tissues and tumors can be dissociated mechanically, such as using a mortar and pestle, or a rotor - stator device. However, these techniques may be limited by inconsistent cell yields, poor cell viability, and user skill.

[0006] Accordingly, there is a need for new methods to dissociate and / or homogenize biological samples such as tissues. A solution to this problem would ideally recover a large number of viable cells. In certain applications, it may be further desirable to dissociate biological samples while minimizing cell homogenization. Summary of the Invention

[0007] The present disclosure relates to devices and methods for processing biological samples such as tissue or tumor samples. More specifically, the present disclosure relates to automated devices and methods for processing biological samples such as tissue or tumor samples.

[0008] In one aspect of the present disclosure, there is provided an apparatus for processing a biological sample. In one embodiment, the apparatus is a cartridge that can be used to process a biological sample. In one embodiment, the cartridge for processing a biological sample may include a lid having a top surface and a bottom; a shaft that is rotatable relative to the lid, the shaft having a first end that mates with the lid and an opposite second end that extends away from the bottom; an agitator attached to the shaft; one or more grooves in an outer peripheral edge of the agitator; a container that can engage the bottom of the lid; and one or more teeth disposed in an inner wall of the container, the one or more teeth protruding toward the interior of the container and passing through the one or more grooves respectively when the agitator rotates about an axis defined by the shaft.

[0009] In one embodiment, the agitator is an impeller. In one embodiment, the agitator has a substantially constant radius along its length (in the direction of the longitudinal axis of the shaft). In one embodiment, the agitator flares radially wider in a direction from the first end toward the second end. In one embodiment, the agitator is integral with the shaft.

[0010] In one embodiment, the cartridge of the present disclosure may further include a plurality of teeth. In one embodiment, the plurality of teeth are arranged annularly in at least a first row of teeth and a second row of teeth. In one embodiment, the first row of teeth protrudes more toward the interior of the container than the second row of teeth. In one embodiment, the first row of teeth is located radially outside the second row of teeth.

[0011] In one embodiment, one or more teeth are disposed in the bottom wall of the container.

[0012] In one embodiment, a cross-section of one or more teeth taken in a plane parallel to the inner wall is an ellipse. In one embodiment, the eccentricity of the ellipse is greater than 0 and less than 1. In one embodiment, the eccentricity of the ellipse is greater than 0.5.

[0013] In one embodiment, the one or more teeth terminate in a tapered edge. In one embodiment, the tapered edge is smooth. In one embodiment, the tapered edge is serrated.

[0014] In one embodiment, the cartridge of the present disclosure may further include alignment features in the bottom wall of the container for engaging the end of the second end of the shaft.

[0015] In one embodiment, the cartridge is a dissociation cartridge for dissociating a tissue sample. In one embodiment, the cartridge is a homogenization cartridge for homogenizing a tissue sample.

[0016] In another aspect of the present disclosure, there is provided an assembled cartridge including one or more of the features described above, wherein the lid is engaged with the container.

[0017] In another aspect of the present disclosure, there is provided a system for processing biological samples. In one embodiment, the system is automated. In one embodiment, the system of the present disclosure may include a base having one or more receiving areas for respectively receiving the assembled cartridges as described herein; at least one rotatable main shaft capable of engaging with the axis of the assembled cartridge, a motor for rotating the main shaft, and at least one processor or microprocessor configured to output a processing protocol or instructions to at least the motor (through a controller or a microcontroller).

[0018] In one embodiment, the system of the present disclosure may further include venting. In one embodiment, the system of the present disclosure may further include a conduit in fluid communication with the inner cavity of the base. In one embodiment, the system of the present disclosure may further include one or more fans for moving air within the system. In one embodiment, the one or more fans are configured to draw air into the conduit and out of the system through the vent.

[0019] In one embodiment, a cartridge (e.g., an assembled cartridge) may be received within the receiving area. In one embodiment, the receiving area includes a hole defined by a hole wall.

[0020] In one embodiment, the system of the present disclosure may further include a Peltier module for establishing or changing the temperature of the hole wall. In one embodiment, the processing protocol includes establishing or changing the temperature of the hole wall.

[0021] In one embodiment, the processing protocol includes setting the speed and / or direction and / or duration of rotation of the motor (and at least one main shaft and associated shaft / stirrer).

[0022] In one embodiment, the processing protocol or instructions are selected or input through a graphical user interface.

[0023] In one embodiment, the system of the present disclosure may further include a sensor downstream of the at least one processor or microprocessor, the sensor relaying a feedback signal to the at least one processor or microprocessor.

[0024] In one embodiment, the main shaft is movable or biasable from a first retracted position to a second extended position.

[0025] In another aspect of the present disclosure, a method for processing a biological sample is provided. The method of the present disclosure may include providing a biological sample in a cartridge (as described herein), performing a processing protocol, and generating a processed biological sample with reduced complexity.

[0026] In one embodiment, performing the processing protocol includes rotating a stirrer to direct the biological sample towards and into contact with one or more teeth disposed in the cartridge. In one embodiment, rotating the stirrer includes rotating for a defined period of time and / or at a defined rotational speed. In one embodiment, rotating the stirrer includes unidirectional rotation. In one embodiment, rotating the stirrer includes reversing the direction of stirrer rotation at least once during the processing protocol. In one embodiment, rotating the stirrer includes oscillating the direction of stirrer rotation during the processing protocol.

[0027] In one embodiment, performing the processing protocol includes incubating at least once. In one embodiment, performing the processing protocol includes incubating more than once. In one embodiment, one or more incubations during the processing protocol are performed at a current temperature ranging between 4°C and 55°C.

[0028] In one embodiment, the biological sample with reduced complexity changes along a continuous process from a shredded state to a dissociated state and then to a homogenized state.

[0029] The method of the present disclosure may further include post-processing the processed biological sample. For example, post-processing may include filtration, washing, myelin removal, DNase treatment, nucleic acid extraction, protein separation, immunostaining, debris removal, cell separation / enrichment, and / or removal / lysis of red blood cells.

[0030] The method of the present disclosure may further include performing further downstream analysis on the processed or post-processed biological sample. For example, further downstream analysis may include: nucleic acid analysis by PCR, qPCR, RT-qPCR, sequencing, or high-throughput sequencing, etc.; protein analysis by Western blotting, immunostaining, proteomics, etc.; or cell analysis by flow cytometry, etc.

[0031] In one embodiment, providing the biological sample in the cartridge (of the present disclosure) may include docking the cartridge with the system as described above. Thus, the method of the present disclosure may be an automated method. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] To better understand the various embodiments described herein and to more clearly show how these various embodiments may be implemented, reference will be made, by way of example, to the accompanying drawings, which show at least one exemplary embodiment and which will now be described. The drawings are not intended in any way to limit the scope of the teachings described herein.

[0033] Figure 1 Shows a perspective front view of the assembled cartridge, where the cap is engaged with the container.

[0034] Figure 2 Shows a perspective front view (A) of the cartridge cap disengaged from the container and a perspective bottom view (B) of the bottom of the cartridge cap disengaged from the container.

[0035] Figure 3 Shows a perspective front view (A) of the cartridge container disengaged from the cap and a top view (B) of the cartridge container disengaged from the cap.

[0036] Figure 4 Shows an exploded perspective view of the cartridge of the present disclosure.

[0037] Figure 5 Shows a perspective view of one embodiment of the system of the present disclosure.

[0038] Figure 6 Shows Figure 5 An enlarged view of the main shaft and receiving area depicted in

[0039] Figure 7 Shows a perspective view of different embodiments of the device of the present disclosure, where a sheath (A) around the main shaft and a close-up (B) of the sheath and the main shaft are depicted.

[0040] Figure 8 Shows bar graphs quantifying the viability and yield of cell suspensions obtained after dissociating spleen ((A) and (B)), brain ((C) and (D)), and lung ((E) and (F)) tissues using the automated system of the present disclosure compared to two commercially available (automated and manual) tissue dissociation systems. Data represent the mean from 2 - 13 experiments. Also shown are bar graphs quantifying the effect of temperature differences during the lung dissociation protocol on the viability (G) and yield (H) of the generated cells. Detailed Description

[0041] The present disclosure relates to systems, devices, and methods for processing biological samples. In one embodiment, the systems, devices, and methods of the present disclosure are automated. In one embodiment, the systems, devices, and methods are for processing biological samples to produce cell suspensions, such as single cell suspensions. In one embodiment, the systems, devices, and methods are for processing biological samples to produce tissue homogenates. In one embodiment, the systems, devices, and methods are for processing biological samples to produce cell suspensions, such as single cell suspensions and / or tissue homogenates.

[0042] The processing of a biological specimen or sample generally results in a reduction in its complexity. In one embodiment, processing a biological specimen or sample results in its dissociation. As used herein, the term "dissociation" refers to reducing the complexity or organization of a biological sample such as tissue or tumor to a plurality of cells. Preferably, most of the dissociated cells are intact and / or alive. Generally, when dissociating a biological sample such as tissue or tumor into a cell suspension, it may be necessary to limit or avoid homogenization of the biological sample (including cells).

[0043] In one embodiment, processing a biological specimen or sample results in its homogenization. In the context used herein, the term "homogenization" refers to reducing the complexity or organization of a biological sample such as tissue or tumor more significantly (relative to dissociation) to a suspension comprising a variety of subcellular components such as organelles, nucleic acids, and other subcellular components. Generally, when homogenizing a biological sample such as tissue or tumor into a suspension of subcellular components, it may be necessary to completely (or substantially completely) rupture the cells, but limit or avoid the breakdown of organelles and other macromolecules.

[0044] The biological samples processed using the systems, devices, and methods of the present disclosure are not particularly limited as long as they contain cells. In one embodiment, the biological sample corresponds to any tissue or any tissue fragment. As non-limiting examples, the tissue can be brain, liver, pancreas, spleen, prostate, lung, or a part thereof. In one embodiment, the biological specimen corresponds to a tumor or a tumor sample.

[0045] After dissociating a biological sample using the disclosed systems, devices, and methods, it may be necessary to use the dissociated cells for downstream experiments, culturing, or assays. In such cases, it is preferred that a substantial portion of the recovered cells are alive. Thus, it is important to process freshly isolated rather than preserved (such as cryopreserved) biological samples using the disclosed systems, devices, and methods. However, not all applications are performed using live cells, such as when cell staining is to be performed, or when subcellular components or macromolecules need to be recovered.

[0046] After homogenizing a biological sample using the disclosed systems, devices, and methods, it may be necessary to use subcellular components such as nucleic acids or proteins for downstream experiments, culturing, or assays. In such cases, it is preferred that a substantial portion of the cells of the tissue have been ruptured. However, it is important to homogenize freshly isolated rather than preserved (such as cryopreserved) biological samples using the disclosed systems, devices, and methods.

[0047] In one embodiment, the disclosed systems, devices, and methods are preferably used to process biological samples such as tissue, tumor, or parts thereof to obtain a cell suspension. Thus, it is advantageous to use systems, devices, and methods that achieve high cell yields, maximize the frequency of live cells, and / or minimize cell homogenization.

[0048] Cartridge

[0049] In one aspect of the present disclosure, a cartridge for processing a biological sample is provided. The cartridges of the present disclosure can be used alone or in combination with the systems of the present disclosure. In one embodiment, the cartridge of the present disclosure docks with an instrument or system of the present disclosure. The cartridge (or the cartridge docked with the system) can be used to perform processing protocols such as chopping, dissociating, or homogenizing protocols on biological samples.

[0050] The cartridges of the present disclosure can be single-use or reusable. In some embodiments, preferably, the cartridge is single-use, such as when the cartridge is used to process biological hazards, or when it is used for a standardized workflow. In some embodiments, preferably, the cartridge is reusable, and in these embodiments the cartridge can be washable and / or disinfectable. In one embodiment, the cartridge of the present disclosure can be selected by the user to be used only once or reused.

[0051] The cartridges of the present disclosure are not particularly limited in terms of one or more manufacturing materials. The cartridge can have sufficient rigidity to withstand reasonable wear. In one embodiment, the cartridge is made of a polymer and can be manufactured by injection molding or 3D printing. In this embodiment, the cartridge can be transparent or translucent to allow the user to observe the progress of biological sample processing without opening the cartridge. In one embodiment, the cartridge is die-cast, such as made of stainless steel.

[0052] The material used to manufacture the cartridge should be able to withstand a temperature range, such as -80°C to 80°C. However, whether at ambient temperature or different temperatures, the material from which the cartridge is made should not leach or decompose when exposed to biological samples or solutions in which biological samples are immersed. In addition, the material used to manufacture the cartridge should not be toxic to the biological samples being processed.

[0053] In one embodiment, the cartridge 1 of the present disclosure can include a plurality of components, such as a lid 5 and a container 7. Referring to Figures 1-4 , the cartridge 1 for processing a biological sample includes a lid 5 having a top surface (exposed to the external environment) and a bottom surface (substantially protected from the external environment when engaged with the container).

[0054] The lid 5 can be fixed to the complementary container 7 in many different ways. In one embodiment, the lid 5 has threads for engaging the container 7, preferably near its opening. In one embodiment, ridges on either the lid or the container can match grooves on the lid or the container, such as by press-fitting.

[0055] The cylinder 1 further includes a shaft 10 having a first end 12 and an opposite second end 14 that define an axis l. The first end 12 mates with the lid 5, and the second end 14 extends away from the bottom of the lid. In one embodiment, the first end 12 is received through a hole 8 in the lid 5. In one embodiment, the first end 12 mates with the bottom of the lid 5.

[0056] The shaft 10 and the lid 5 are rotatable relative to each other. In one embodiment, the axis of rotation is defined by the longitudinal axis l of the shaft 10 (e.g., the axis defined by passing through the first end 12 and the opposite second end 14). Although the shaft and the lid are rotatable relative to each other, it is important that the shaft 10 and the lid 5 form a leak-proof seal. In one embodiment, the shaft 10 (and / or the first end 12) and the lid 5 are in a sealed (or leak-proof) engagement, yet still allow the shaft 10 to rotate within the hole 8. The ways of sealing relatively movable components are known and may include grommets, gaskets, bearings, etc. (as shown by the seal 13 in Figure 4 .

[0057] The cylinder 1 further includes an agitator 15 attached to the shaft 10. The agitator 15 can be a separate component connected to the shaft 10 or integrally formed with the shaft 10. The agitator 15 rotates within the cylinder 1. In one embodiment, the agitator 15 rotates independently about the axis l (as defined by the shaft 10). In one embodiment, the agitator 15 rotates together with the shaft 10 about the axis l (as defined by the shaft 10).

[0058] The agitator 15 can take any form as long as it can direct the movement of the fluid or substance (such as a biological sample or a fragment of a biological sample) with which it comes into contact. In one embodiment, the agitator 15 can both move the substance with which it comes into contact and direct the substance in a specific direction (e.g., downward toward the bottom wall of the container or through a cutting surface). In one embodiment, the agitator 15 is an impeller. In one embodiment, the agitator 15 is a fin. In such an embodiment, the agitator 15 can flare outwards from the shaft 10. In one embodiment, the agitator 15 flares out laterally from the shaft 10.

[0059] In one embodiment, the agitator 15 includes a side edge or an outward edge 17 (see Figure 2 and Figure 4 ). The side edge 17 refers to the part of the agitator 15 that is farthest from the shaft 10 as measured in a plane orthogonal to the shaft 10, more specifically, to the axis l.

[0060] In one embodiment, the agitator 15, more specifically, the side edge 17, flares out wider radially in the direction from the first end 12 to the second end 14.

[0061] In one embodiment, the distance r of the side edge 17 from the shaft 10 or the axis l (taken in a plane orthogonal to the axis l) is constant or substantially constant along the length of the agitator 15 (e.g., along the axis l defined by the first end 12 and the second end 14). The constancy or substantial constancy of the side edge 17 may be important for minimizing the gap between the side edge 17 (of the agitator 15) and the inner (side) wall of the container 7, thereby reducing or limiting the spillage of the biological sample upon contact with the agitator 15.

[0062] The agitator 15 may further include one or more grooves 20 (which may be successively referred to as 20a, 20b, etc.) in its outer peripheral edge 22 (see Figure 2 ). In one embodiment, the outer peripheral edge 22 extends along a plane orthogonal to the axis l. In one embodiment, the plane in which the outer peripheral edge 22 extends (along the distance r) is in the same plane as the plane orthogonal to the end of the second end 14. In one embodiment, the plane in which the outer peripheral edge 22 extends is parallel to the plane orthogonal to the end of the second end 14. In one embodiment, the plane in which the outer peripheral edge 22 extends is lower than the plane orthogonal to the end of the second end 14, that is, the outer peripheral edge 22 extends further from the first end 12 than the end of the second end 14. In one embodiment, the plane in which the outer peripheral edge 22 extends is higher than the plane orthogonal to the end of the second end 14, that is, the end of the second end 14 extends further from the first end 12 than the outer peripheral edge 22. In one embodiment, the outer peripheral edge 22 is the same edge as the side edge 17.

[0063] Referring to Figure 3 , the cartridge 1 for processing a biological sample further includes a container 7 capable of engaging the bottom of the lid 5. The container 7 can be made of any material as long as it can hold liquids and solids. In one embodiment, the container 7 can be made of the same material as the lid 5. In one embodiment, the container 7 can be made of a material different from the material used to make the lid 5. Whether the lid 5 and the container 7 are made of the same or different materials, it is important that the two components are capable of mating, connecting, or attaching.

[0064] In one embodiment, the lid 5 and the container 7 are sealed when engaged or mated with each other.

[0065] The container 7 may include one or more teeth 33 (which may be successively referred to as 33a, 33b, etc.) disposed in its inner wall 35. In one embodiment, the container 7 includes a plurality of teeth disposed in its inner wall 35. In one embodiment, the container 7 includes a single tooth disposed in its inner wall 35.

[0066] In one embodiment, the inner wall 35 is the bottom wall of the container 7. In one embodiment, the inner wall 35 is the side wall of the container 7. The wall of the container 7 in which one or more teeth 33 are arranged will depend on the position of one or more slots 20 configured on the agitator 15. If one or more slots 20 are configured in the side edge 17, one or more teeth 33 are arranged in the side wall of the container 7, particularly in the inner side wall of the container 7. If one or more slots 20 are configured in the outer peripheral edge 22, one or more teeth 33 are arranged in the bottom wall of the container 7, particularly in the inner bottom wall of the container 7.

[0067] One or more teeth 33 may project towards the interior of the container 7. In one embodiment, one or more teeth 33 project orthogonally to the inner wall 35 and towards the interior of the container 7. In one embodiment, one or more teeth 33 project towards the interior of the container 7 at an angle that is not orthogonal to the inner wall 35.

[0068] As described above, the angle at which one or more teeth 33 project towards the interior of the container 7, whether orthogonal or otherwise, is not particularly limited. However, importantly, regardless of the angle at which one or more teeth 33 project from the inner wall 35, they will pass through one or more slots 20, such as when the shaft 10 (and / or the agitator 15) rotates about the axis l.

[0069] In one embodiment, a plurality of teeth 33 are arranged annularly in the inner wall 35 of the container 7, such as in its bottom wall. In one embodiment, a plurality of teeth 33 are arranged annularly or concentrically in at least a first row of teeth 37 and a second row of teeth 39. In one embodiment, the first row of teeth 37 is located radially outside the second row of teeth 39. In one embodiment, the first row of teeth 37 is located radially inside the second row of teeth 39. If the container 7 includes more than two rows of annularly or concentrically arranged teeth, these additional rows of teeth may be positioned sequentially relative to the first row of teeth 37 and the second row of teeth 39.

[0070] In one embodiment, a plurality of teeth 33 are arranged circumferentially in the inner wall 35 of the container 7, such as in its side wall (not shown). In one embodiment, a plurality of teeth 33 are arranged circumferentially in at least a first row (e.g., row) of teeth 37 and a second row (e.g., row) of teeth 39. In one embodiment, the first row (e.g., row) of teeth 37 is positioned closer to the opening of the container 7 than the second row (e.g., row) of teeth 39. In one embodiment, the first row (e.g., row) of teeth 37 is positioned farther from the opening of the container 7 than the second row (e.g., row) of teeth 39. If the container 7 includes more than two rows (e.g., rows) of circumferentially arranged teeth, these additional rows (e.g., rows) of teeth may be positioned sequentially relative to the first row (e.g., row) of teeth 37 and the second row (e.g., row) of teeth 39.

[0071] In one embodiment, the first row of teeth 37 and the second row of teeth 39 project into the interior of the container 7 to the same or substantially the same extent. In one embodiment, the first row of teeth 37 extends more into the interior of the container 7 than the second row of teeth 39. In one embodiment, the second row of teeth 39 extends more into the interior of the container 7 than the first row of teeth 37.

[0072] The shape of the one or more teeth 33 is not particularly limited, provided that their dimensions are designed to pass (respectively) through the one or more slots 20, such as when the shaft 10 (and / or the agitator 15) rotates about the axis l. The following description of exemplary tooth shapes is made in a plane parallel to the inner wall in which they are arranged.

[0073] In one embodiment, the cross-section of the one or more teeth 33 (or the plurality of teeth) is elliptical. In these embodiments, the eccentricity of the ellipse is greater than 0 and less than 1. In one embodiment, the eccentricity of the ellipse is about 0.5 or greater (and less than 1).

[0074] In one embodiment, the cross-section of the one or more teeth 33 (or the plurality of teeth) is curved or arcuate. In one embodiment, the curved or arcuate cross-sectional shape can correspond to the radius of curvature of the ring in which the one or more teeth 33 can be arranged.

[0075] In one embodiment, the cross-section of the one or more teeth 33 (or the plurality of teeth) is arcuate or annular or oval. In one embodiment, the cross-section of the one or more teeth 33 (or the plurality of teeth) is a different polygon, such as a quadrilateral, pentagon, hexagon, rhombus, etc. In this embodiment, as in all embodiments, the only constraint on the shape and height of the teeth is that they pass through the one or more slots 20, respectively, such as when the shaft 10 (and / or the agitator 15) rotates about the axis l.

[0076] The processing of biological samples, such as dissociation or homogenization, can be facilitated by design features included on the plurality of teeth 33. In one embodiment, one or more of the teeth 33 terminate in or include a tapered edge 40. The tapered edge 40 can facilitate the breakdown of the biological sample contained in the container 7, particularly when the agitator 15 moves the biological sample near the plurality of teeth 33 while rotating about the axis l. In one embodiment, the tapered edge 40 is smooth. In one embodiment, the tapered edge 40 is serrated. In one embodiment, a subset of the plurality of teeth 33 is smooth and the remaining teeth are serrated.

[0077] The container 7 may further include alignment features 42 for engaging the second end 14 (or its terminus) of the shaft 10. When the second end 14 of the shaft 10 engages the alignment features 42, it can ensure and maintain the agitator 15 rotating about a consistent (non-offset) axis of rotation l while limiting or avoiding incorrect alignment of one or more teeth 33 and one or more slots 20.

[0078] In one embodiment, the alignment features 42 are positioned or configured in the inner wall 35 of the container 7. In one embodiment, the alignment features 42 are positioned or configured in the bottom wall of the container 7. In such an embodiment, the alignment features 42 may be a pit or depression in the bottom wall. In one embodiment, the alignment features 42 are positioned or configured in the side wall of the container 7. In such an embodiment, the alignment features 42 may be a circumferential groove in the side wall.

[0079] In the case where the cartridge 1 is used to dissociate a biological sample such as tissue or a tumor, the cartridge may be regarded as a dissociation cartridge. In the case where the cartridge 1 is used to homogenize a biological sample such as tissue or a tumor, the cartridge may be regarded as a homogenization cartridge.

[0080] In one aspect, the cartridge 1 may be provided in component parts (e.g., a lid 5 separate from the container 7). In one aspect, the cartridge 1 may be provided together or in an assembled form (e.g., the lid 5 engaged with the container 7).

[0081] System

[0082] In one aspect of the present disclosure, a system for processing a biological specimen or sample is provided. In one embodiment, the system is automated. In one embodiment, the system docks with a cartridge as described above.

[0083] The system 100 (in combination with the cartridge 1) can be used to dissociate and / or homogenize a biological sample, such as a tissue or tumor sample.

[0084] Referring Figures 5 to 7 , the system 100 may include a base 102 and a head 104 connected to the base by a rear wall 106.

[0085] The base 102 may include one or more receiving areas 110 for receiving cartridges 1 (e.g., assembled cartridges 1, more specifically their containers 7) respectively. In one embodiment, the base 102 includes a plurality of receiving areas 110 for receiving cartridges 1 (e.g., assembled cartridges 1, more specifically their containers 7) respectively. In one embodiment, the base 102 includes 2 receiving areas 110, 3 receiving areas 110, 4 receiving areas 110, 5 receiving areas 110, 6 receiving areas 110, 7 receiving areas 110, 8 receiving areas 110, 9 receiving areas 110, 10 receiving areas 110, 11 receiving areas 110, 12 receiving areas 110 or more.

[0086] Each receiving area 110 may include a hole 115 defined by a hole wall 120. The cross-sectional shape of the hole 115 is not particularly limited as long as it accommodates the container 7 and avoids or restricts the swinging or lateral movement of the container 7 within the hole 115 (such as by a hole 115 that is deep enough relative to the height of the container 7). In some embodiments, the cross-sectional shape of the hole 115 is the same as or substantially the same as the cross-sectional shape of the container 7 (both taken in a plane defined by 0 degrees and 180 degrees). In some embodiments, the cross-sectional shape of the hole 115 is different from or larger than the cross-sectional shape of the container 7. In such embodiments, it is important to use an adapter or configure guiding features within the hole 115 to control the swinging or lateral movement of the container 7 therein.

[0087] The size of the hole 115 may have a diameter or width that is wider / larger than the diameter of the container 7 of the cartridge 1 by about 0.1 mm, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1.5 mm, about 2 mm, about 2.5 mm. In one embodiment, the size of the hole 115 may have a diameter or width that is wider / larger than the diameter of the container 7 of the cartridge 1 by about 2.5 mm.

[0088] In one embodiment of the system 100, one or more receiving areas 110 are in fixed positions within the base 102. In one embodiment of the system 100, one or more receiving areas 110 are movable (e.g., rotatable) within the base 102, such as on a turntable integrated into or on the base 102.

[0089] The base 102 may further include one or more Peltier modules 125. In one embodiment, each hole 115 is respectively associated with a Peltier module 125. Thus, in one embodiment, the system 100 includes the same number of Peltier modules as the receiving area 110 (and holes 115). Although the following description may apply to each of one or more Peltier modules 125, for simplicity, only a single Peltier module 125 will be referred to.

[0090] In one embodiment, the Peltier module 125 is integrated into the base 102 and establishes or regulates the temperature of at least a portion of the base 102, such as the hole wall 120. In one embodiment, the Peltier module 125 is communicatively coupled to at least one processor or microprocessor. Thus, under a sample processing scheme selected or input by the user, the temperature of the base 102, and more particularly, the temperature of the receiving area 110, and more particularly the temperature of the hole wall 120, can be controlled.

[0091] The system 100 further includes one or more spindles 150 connected to the head 104. One or more spindles may engage or be capable of engaging with the axis 10 of the cartridge 1 (when the cartridge 1 is located in the receiving area 110 and / or hole 115), and more particularly, with the first end 12 of the axis. In one embodiment, one or more spindles 150 hang down from the head 104. In one embodiment, the number of spindles corresponds to the number of receiving areas 110, and more particularly, to the number of holes 115.

[0092] In one embodiment, one or more spindles 150 are rotatable. Although the following description may apply to each of one or more spindles 150, for simplicity, only a single spindle 150 will be referred to. In one embodiment, each spindle 150 rotates about the same axis l as the axis 10 of the cartridge 1.

[0093] In embodiments of the system 100 that include rotatable spindles, the system 100 includes one or more motors 170, such as stepper motors. Generally, one or more motors 170 are located within the head 104. In one embodiment, a corresponding motor transfers rotational motion to a corresponding spindle 150. In one embodiment, a single motor transfers rotational motion to multiple spindles 150.

[0094] In one embodiment, the spindle 150 itself is not rotatable, but includes a transmission member that transfers rotational motion to the axis 10 (or agitator 15). In one embodiment, the spindle 150 itself is not rotatable, but includes wiring that communicates with a rotor assembly that transfers rotational motion to the axis 10 (or agitator 15).

[0095] The main shaft 150 can be movable or deflectable between a first retracted position and a second extended position. Thus, the main shaft 150 can cooperate with the receiving area 110 (and the hole 115) to accommodate cartridges 1 of different heights and / or different shaft 10 lengths.

[0096] The direct or indirect engagement or mating of the main shaft 150 with the shaft 10 (more particularly, with the end 12) may cause the agitator 15 to rotate under the influence of a motor such as motor 170. The manner in which the main shaft 150 and the shaft 10 are engaged or mated is not particularly limited. In some embodiments, the engagement or mating of the main shaft 150 and the shaft 10 is achieved through complementary male and female connections. The male connector can be positioned on the main shaft 150 and the female connector can be positioned on the shaft 10 (or more particularly positioned on the end 112), and vice versa.

[0097] In any case, the male and female connection mating parts are formed in a complementary mating manner and are formed in a manner to avoid or limit sliding between them, such as when the main shaft 150 rotates. In one embodiment, when viewed in a plane orthogonal to the longitudinal axis l of the shaft, the male connector includes at least one straight edge that can mate with a corresponding shaped female connector mating part. In one embodiment, the male connector includes more than one straight edge that can mate with a corresponding shaped female connector mating part. As a non-limiting example, the male connector can be semi-circular ring-shaped, triangular, square, pentagonal, hexagonal or any other shape including at least one straight edge. When torque is applied to it, a circular or arc-shaped shape may slide or eventually slide, but a connection mediated by at least one straight edge or more than one straight edge can better withstand higher levels of torque without causing relative sliding of the two connectors.

[0098] In a specific embodiment, the male connector including at least one, or more than one straight edge (when viewed in a plane orthogonal to the longitudinal axis of the shaft 10) projects from the shaft 10 (or the end 12) in the direction of the main shaft 150, and the main shaft 150 includes a female connector that can mate with the male connector (e.g., complementary mating with it) (see Figure 4 and Figure 5 ).

[0099] In a specific embodiment, the shaft 10 (or in the end 12) is configured with a female connector including at least one or more than one straight edge (when viewed in a plane orthogonal to the longitudinal axis of the shaft 10), and the main shaft 150 includes a male connector that projects in the direction of the shaft 10 (or the end 112), and the male connector can mate with the female connector (e.g., complementary mating with it).

[0100] The system 100 may further include a sheath 175 that surrounds or at least partially surrounds the main shaft 150 (Figure 7 )。Similar to the main shaft 150, the sheath 175 can be movable or deflectable between a first retracted position and a second extended position. The sheath 175 can be fully retracted into the head 104, or only partially retracted, and the sheath 175 can extend towards or into the base 102. In one embodiment, the main shaft 150 and the associated sheath 175 are movable relative to each other (along axis l).

[0101] In one embodiment, the main shaft 150 and the associated sheath 175 are movable relative to each other (along axis l), but only along a defined path until the engagement feature of the sheath 175 contacts an upper or lower limit (not shown) of the main shaft 150. By further description, when the sheath 175 moves upward to the retracted position, the engagement feature on the inward-facing surface of the sheath 175 contacts the upper limit on the main shaft 150, causing the main shaft and the sheath 175 to retract simultaneously. Also, when the sheath 175 moves downward to the extended position, the engagement feature on the inner surface of the sheath 175 contacts the lower limit on the main shaft 150, causing the main shaft and the sheath 175 to extend simultaneously. Thus, a single upward or downward movement of the sheath 175 along axis l can extend or retract both the sheath 175 and the associated main shaft 150.

[0102] In embodiments where the system 100 is automated, a large amount of heat may be generated during operation, especially when one or more Peltier modules 125 and one or more motors 170 are both operating. To mitigate the heat buildup to a high or unsafe level, vents can be provided in the system 100, such as in the base 102 and / or the head 104 and / or the rear wall 106 and / or the side panels. The vents can include a single vent hole or multiple vent holes / perforations. In one embodiment, vents / perforations can be provided in the bottom wall of the base 102. In one embodiment, vents / perforations can be provided in one or more side walls of the base 102. In one embodiment, vents / perforations can be provided in both the bottom wall and one or more side walls of the base 102. In one embodiment, vents / perforations can be provided in the rear wall 106 (or panel) of the system 100. In one embodiment, vents / perforations can be provided in the head 104, which substantially overlap the location where one or more motors 170 are located.

[0103] In one embodiment, the vent can be provided as a slit in the head 104, which substantially overlaps the location where one or more motors 170 are located. In the same or different embodiments, the vent or the component of the vent can be provided as a slit or perforation in the base 102, which substantially overlaps the location where one or more Peltier modules 125 are located.

[0104] System 100 may further include internal pipes (not shown). In one embodiment, the pipes are in fluid communication with the inner cavity of the base 102.

[0105] Vents and / or pipes may cooperate with other cooling features of the system 100, such as one or more fans, to bring external air into the system 100 and remove the heated air from the system 100. In one embodiment, the system 100 may include a fan configured to draw external air into the system 100 and pass it through and out of the system 100 via the vents. In one embodiment, the system 100 includes a series of fans within the base 102 and / or the head 104 and / or the rear wall 106.

[0106] Thus, the cooperation of one or more fans and vents (such as slits and / or perforations) reduces heat accumulation to ensure the safe operation of the various thermoelectric modules of the system 100 and that they do not overheat.

[0107] In an automated embodiment of the system 100, it includes at least one processor or microprocessor. The at least one processor may be configured to output a processing scheme to at least one controller or microcontroller. In one embodiment, one or more processing schemes may be pre-programmed into the system 100. In one embodiment, one or more processing schemes may be input into the system 100 by a user. In one embodiment, the system 100 may include one or more pre-programmed processing schemes and customizable and / or inputtable processing schemes.

[0108] The user may select or input one or more processing schemes through the graphical user interface 300. In one embodiment, one or more processing schemes may include, in any order: one or more incubation steps; one or more temperature adjustments; and one or more agitation steps. Thus, the processing scheme may define the steps for dissociating or homogenizing a biological sample.

[0109] The graphical user interface 300 may be included within the system 100 or may be external to the system 100. In one embodiment, the graphical user interface 300 is connected to the Internet or a mobile communication network. In this embodiment, the user may remotely select or input a processing scheme, such as via a mobile device application. The graphical user interface may be a computer or a tablet and thus includes at least one processor or microprocessor communicatively coupled to at least one controller or microcontroller (which guides the operation of at least one or more motors 170 and one or more Peltier modules 125).

[0110] The processing solution may include setting or adjusting one or more of the following: the rotational speed of the motor 170 (and the corresponding agitator 15); the rotation duration of the motor 170 (and the corresponding agitator 15); and the rotation direction of the motor 170 (and the corresponding agitator 15). In one embodiment, depending on the stage of the solution, the processing solution includes setting or adjusting two or each of the rotational speed of the motor 170 (and the corresponding agitator 15), the rotation duration of the motor 170 (and the corresponding agitator 15), and the rotation direction of the motor 170 (and the corresponding agitator 15).

[0111] In some embodiments, the processing solution may include establishing or changing the temperature of the pore wall 120. Establishing or changing the temperature of the pore wall 120 may be important when the container contains an enzyme solution to facilitate processing of a biological sample therein and the enzyme solution works optimally at a specific temperature or temperature range. Thus, in such cases, it may be important that the container 7 is in close contact with the pore wall 120 and the container 7 is made of a material that readily conducts or radiates an externally applied temperature. Establishing or changing the temperature of the pore wall 120 may also be important when the biological sample being processed needs to be incubated at a specific temperature, such as incubated at about 4 °C to preserve the processed sample before the user collects it, and / or incubated at about 37 °C to obtain optimal enzyme activity.

[0112] In operation, the dissociation or homogenization or incubation solution includes sequential individual steps, which may include one or more incubations (at one or more predetermined temperatures) and one or more agitation steps (at one or more predetermined agitation rates and / or directions and / or durations).

[0113] The system 100 may further include at least one sensor downstream of at least one processor or microprocessor to relay a feedback signal to the at least one processor or microprocessor so that corrective action can be taken. Exemplary signals include but are not limited to the temperature inside the system 100, the temperature of the pore wall 120, the mass load on or in the receiving area 110 and / or the pores 115, or the proper connection between the spindle 150 and the shaft 10.

[0114] Regardless of the feedback signal, at least one processor or microprocessor can be programmed to take corrective action or abort the processing scenario. For example, if the temperature of system 100 and / or the hole wall 120 is outside the acceptable range, the processing scenario will not be executed until the user intervenes or the temperature falls within the acceptable range. As another example, if the mass load on and / or in the receiving area 110 and / or the hole 115 is outside the acceptable range, the processing scenario will not be executed until the user intervenes or the mass load falls within the acceptable range. As another example, if the sheath 175 is not fully extended, the processing scenario will not be executed until the user intervenes or the mass load falls within the acceptable range. As another example, if the spindle 150 is not properly engaged with the shaft 10, the processing scenario will not be executed until the user intervenes or the mass load falls within the acceptable range.

[0115] System 100 can be modular in that multiple systems can be interconnected or daisy-chained to increase or decrease the number of receiving areas for the cartridge 1. Similarly, system 100 can be modular in that the number of receiving areas of a single system can be increased or decreased.

[0116] In one embodiment, the primary system (as described above) includes at least one processor (or microprocessor) and at least one controller (or microcontroller), and the secondary system daisy-chained thereto also includes at least one processor (or microprocessor) and at least one controller (or microcontroller).

[0117] In one embodiment, the primary system (as described above) includes at least one processor (or microprocessor) and at least one controller (or microcontroller), and the secondary system daisy-chained thereto does not include at least one processor (or microprocessor). Instead, the secondary system can rely on at least one processor (or microprocessor) of the primary system to communicate the processing scenario to it (and more specifically, to its at least one controller (or microcontroller)). Thus, the processing scenarios for each of the primary and secondary systems can be selected or input through a single interface such as interface 300.

[0118] System 100 (whether primary, secondary, etc.) can include a secondary display 310 to provide feedback to the user or cross-reference certain information such as system or spindle / receiving area identifiers.

[0119] In one embodiment, whether only the primary system is deployed or the primary and secondary systems are connected to each other, the user can execute different processing scenarios on each system. In one embodiment, whether only the primary system is deployed or the primary and secondary systems are connected to each other, the user can execute different processing scenarios on each cartridge placed in the receiving area of the corresponding system.

[0120] Method

[0121] In one aspect of the present disclosure, a method for processing a biological sample is provided. The biological sample can be processed, dissociated, and / or homogenized using a cartridge (as described above) and / or a system (also as described above). In one embodiment, the biological sample is processed, dissociated, and / or homogenized using a cartridge (as shown above) docked to a system (as described above).

[0122] The methods of the present disclosure can be performed on any biological sample that includes cells. In one embodiment, the biological sample is an organ or a part of an organ. In one embodiment, the biological sample is a tumor or a part thereof. However, the types of biological samples that can be processed, dissociated, and / or homogenized using the cartridges and / or systems of the present disclosure are not necessarily limited to samples obtained from a subject or patient. For example, the biological sample can be a soil sample that includes single-celled and multi-celled organisms, or a sample of plant material.

[0123] Providing the biological sample in the cartridge (as described above) can encompass suspending the biological sample in a suitable solution. The solution for suspending the biological sample is not particularly limited, but in some embodiments, it is preferred that such a solution is non-toxic to the biological sample (and the resulting cell suspension or homogenate). In one embodiment, the solution for suspending the biological sample is substantially isotonic with the cells, cell organelles, or other membrane-found components contained in the biological sample.

[0124] The biological sample can be suspended or incubated in a solution that includes one or more enzymes. Enzymes for decomposing tissues and tumors (e.g., dissociating and / or homogenizing) are well-known and commercially available. The one or more enzymes included in the solution can be selected from the group consisting of serine proteases, cysteine proteases, aspartic proteases, and metalloproteases. Additionally or alternatively, the one or more enzymes included in the solution can include one or more of the following: trypsin, chymotrypsin, elastase, dispase, thermolysin, collagenase, hyaluronidase, papain, calpain, lysosomal cathepsin, pepsin, subtilisin, rennin, and carboxypeptidase.

[0125] In one embodiment, the enzyme can include an enzyme that has catalytic activity at a temperature above 25°C. In one embodiment, the enzyme can include an enzyme that has catalytic activity at a temperature below 25°C. In one embodiment, the enzyme can include a cold-active enzyme produced by a cold-adapted microorganism. In one embodiment, the cold-active enzyme can include a protease.

[0126] If the biological sample to be processed comprises a dissociative suspension of (intact and / or live) cells, careful selection of the enzyme and its concentration may be important to avoid digesting or cleaving, for example, proteins or glycoproteins associated with the cell membrane.

[0127] As described above, a rotary or oscillating agitator can bring the biological sample into contact with one or more teeth of the consumable, thereby reducing the complexity of the biological sample. Depending on the rate, duration, and direction of rotation or oscillation of the shaft (and agitator), the reduction in the complexity of the biological sample can range from chopping to dissociation to homogenization.

[0128] In one embodiment, rotation or oscillation of the shaft (and agitator) is achieved in the system (as described above) by (biasingly) engaging / matching one or more main shafts with the corresponding shaft ends of the cartridge. In one embodiment, the rotation or oscillation of the agitator is controlled by a (micro)processor, a (micro)controller, and a motor.

[0129] In one embodiment, using the cartridge and system of the present disclosure to dissociate and / or process and / or homogenize a biological sample may require the user to select or input a protocol, such as via a graphical user interface integrated with the system or via a mobile application.

[0130] In one embodiment, after selecting an appropriate protocol, these methods can produce a processed biological sample in which the dissociated cells exhibit a high degree of viability, such as >70%, >75%, >80%, >85%, >90%, or >95% viable cells.

[0131] In one embodiment, after selecting an appropriate protocol, these methods can produce a processed biological sample in which a high number of cells are recovered, such as per milligram of processed tissue. In one embodiment, >1000 cells are recovered per milligram of processed tissue. In one embodiment, >5000 cells are recovered per milligram of processed tissue. In one embodiment, >10000 cells are recovered per milligram of processed tissue. In one embodiment, >15000 cells are recovered per milligram of processed tissue. In one embodiment, >20000 cells are recovered per milligram of processed tissue. In one embodiment, >25000 cells are recovered per milligram of processed tissue. In one embodiment, >30000 cells are recovered per milligram of processed tissue.

[0132] In one embodiment, after selecting an appropriate protocol (e.g., dissociation protocol), these methods can produce a processed biological sample in which a low degree of homogenization occurs. In other words, in the dissociated sample, a small number of cells are broken down and a large number of intact cells are present. The degree of homogenization can be determined by analyzing the amount of one or more macromolecules in the processed sample, such as intracellular proteins, cell membrane fragments, or genomic DNA.

[0133] In one embodiment, compared to other ways of processing biological samples (e.g., manual processing by mortar and pestle, or using different automated means), the methods using the cartridges and / or systems of the present disclosure can produce a comparable or improved yield of viable cells and / or recovered cells (such as per milligram of tissue).

[0134] The biological sample processing performance of an automated system docked with a cartridge (both as described herein and Figures 1 to 7 as depicted) was evaluated. Spleen, lung, and brain samples collected from C57BL / 6 mice were separately placed in containers (as described above), placed in a suitable solution and capped. The assembled cartridge was placed in the receiving area of the automated system of the present disclosure, and a processing protocol (e.g., dissociation protocol) was selected.

[0135] In addition, post-processing may be important in certain applications or for certain tissue or tumor types, such as to further clarify the processed sample. In one embodiment, additional DNase treatment may be important for reducing the viscosity of the processed sample. In one embodiment, the processed sample can be passed through a filter to remove relatively (relative to the filter pore size) large particles and debris.

[0136] Further downstream assays or analyses can be performed on the dissociated cell suspension. For example, cell separation or enrichment assays can be performed on the dissociated cell suspension using conventional methods such as density gradient centrifugation, RBC lysis, or immunomagnetic separation. As another example, the dissociated cell suspension can be characterized by flow cytometry to assess viability and yield ( Figure 8 ). As Figure 8 can be seen, the methods of the present disclosure can be comparable to or superior to other commercially available choices for dissociating samples.

[0137] For spleen samples processed by the methods using the systems and cartridges of the present disclosure, more than 90% of the dissociated cells are viable, and the viability observed in commercially available automated systems is slightly lower. In fact, significantly lower cell viability was observed in manually dissociated spleen sample cells ( Figure 8 A). In terms of cell yield, significantly higher levels were obtained using the methods and systems of the present disclosure compared to the two commercially available methods tested ( Figure 8 B).

[0138] For brain samples processed by the methods using the systems and cartridges of the present disclosure, more than 80% of the dissociated cells are viable, with slightly lower viability observed in the cells dissociated using the two commercially available methods tested ( Figure 8 C). In terms of cell yield, significantly higher levels were obtained using the methods and systems of the present disclosure compared to the two commercially available methods tested ( Figure 8 D).

[0139] For lung samples processed by the methods using the systems and cartridges of the present disclosure, nearly 80% of the dissociated cells are viable, with the same or slightly lower viability observed in the cells dissociated using the two commercially available methods tested ( Figure 8 E). In terms of cell yield, higher levels were obtained using the methods and systems of the present disclosure compared to the two commercially available methods tested ( Figure 8 F).

[0140] The effect of temperature difference during the lung dissociation protocol was also studied. The results showed that while viability was slightly negatively affected when performing the dissociation protocol at 37°C compared to 20°C ( Figure 8 G), the yield of viable cells was significantly higher when performing at 37°C compared to 20°C ( Figure 8 H).

[0141] Thus, compared to commercially available tissue dissociation systems, the automated system of the subject exhibits superior performance, with higher cell yields for both brain and spleen tissues.

[0142] Glossary of Terms

[0143] Although the processes, steps, or blocks are presented in a given order, alternative instances can execute routines with steps in a different order, or employ systems with blocks or steps, and some processes or steps and blocks can be deleted, moved, added, subdivided, combined, and / or modified to provide alternatives or sub - combinations. Each of these processes or steps or blocks can be implemented in many different ways. Additionally, although the processes or steps or blocks are sometimes shown as being executed serially, these processes or steps or blocks can also be executed in parallel, or at different times.

[0144] Furthermore, although elements are sometimes shown as being executed in sequence, they can also be executed simultaneously or in a different order. Accordingly, it is intended that the following claims be construed to include all such variations within their intended scope.

[0145] In the case of referring to a certain component as mentioned above, unless otherwise specified, the reference to the component (including the reference to "way") shall be construed to include any equivalent component that performs the function of the said component (i.e., functionally equivalent), including those components that are not structurally equivalent to the disclosed structure that performs the function in the exemplary embodiments shown in the present invention.

[0146] Various features are described herein as being present in "one embodiment" or some "some embodiments". These features are not essential and may not be present in all embodiments. Embodiments of the present invention may include any combination of zero, any one, or two or more such features. This is only limited to a certain extent, that is, when some such features are incompatible with other such features, specifically, it is impossible for a person of ordinary skill in the art to construct a practical embodiment that combines these incompatible features. Therefore, the description that "some embodiments" have feature A and "some embodiments" have feature B should be construed to clearly indicate that the inventor has also considered embodiments that combine feature A and B (unless the description otherwise states or feature A and B are fundamentally incompatible).

[0147] Therefore, it is intended that the appended claims and the claims introduced hereinafter be construed to include all such reasonably inferable modifications, permutations, additions, omissions, and sub - combinations. The scope of the claims should not be limited by the preferred embodiments set forth in the examples, but should be given the broadest interpretation consistent with the entire specification.

Claims

1. A cartridge for processing a biological sample, comprising a lid having a top surface and a bottom; a shaft rotatable relative to the lid, the shaft having a first end mating with the lid and an opposite second end extending away from the bottom; a stirrer attached to the shaft; one or more grooves in the outer peripheral edge of the stirrer; a container engageable with the bottom of the lid; and one or more teeth disposed in the inner wall of the container, the one or more teeth protruding towards the interior of the container and passing through the one or more grooves when the stirrer rotates about an axis defined by the shaft.

2. The cartridge according to claim 1 or any other claim herein, wherein, The stirrer is an impeller.

3. The cylinder according to claim 1 or 2 or any other claim herein, wherein, The stirrer has a substantially constant radius.

4. The cartridge according to any one of claims 1 to 3 or any other claim herein, further comprising a plurality of teeth.

5. A cartridge according to claim 4 or any other claim herein, wherein, The plurality of teeth are radially arranged in at least a first row of teeth.

6. The cartridge according to claim 5 or any other claim herein, further comprising a second row of teeth concentric with the first row of teeth, wherein, The first row of teeth is radially outside the second row of teeth.

7. A cylinder according to claim 6 or any other claim herein, wherein, The first row of teeth protrudes more towards the interior of the container than the second row of teeth.

8. The cartridge according to any one of claims 1 to 7 or any other claim herein, wherein, The one or more teeth are disposed in the bottom wall of the container.

9. The cartridge according to any one of claims 1 to 8 or any other claim herein, wherein, The cross-section of the one or more teeth taken in a plane parallel to the inner wall is an ellipse.

10. A cylinder according to claim 9 or any other claim herein, wherein, The eccentricity of the ellipse is greater than 0 and less than 1, preferably greater than 0.

5.

11. The tube according to any one of claims 1 to 10, wherein, The one or more teeth terminate in a tapered edge.

12. The cartridge according to claim 11 or any other claim herein, wherein, The tapered edge is smooth or serrated.

13. The cartridge according to claim 8 or any other claim herein, further comprising alignment features in the bottom wall of the container for engaging the end of the second end of the shaft.

14. A cylinder according to any one of claims 1 to 13 or any other claim herein, wherein, The cartridge is a dissociation cartridge and / or a homogenization cartridge for dissociating and / or homogenizing a tissue sample.

15. An assembled cartridge according to any one of claims 1 to 14 or any other claim herein, wherein, The lid engages with the container.

16. An automated system for processing a biological sample, comprising: a base having one or more receiving areas for respectively receiving the assembled cartridge according to claim 15; at least one rotatable main shaft engageable with the shaft of the assembled cartridge; a motor for rotating the main shaft; and at least one processor or microprocessor configured to output a processing protocol to at least the motor.

17. The system according to claim 16 or any other claim herein, wherein, The at least one rotatable main shaft engages with the shaft of the assembled cartridge through complementary and mating connections.

18. The system according to claim 16 or 17 or any other claim herein, wherein, The assembled cartridge is received in a receiving area including a hole defined by a hole wall.

19. The system according to claim 18 or any other claim herein, further comprising a Peltier module for establishing or changing the temperature of the hole wall.

20. The system according to claim 19 or any other claim herein, wherein, The processing protocol includes establishing or changing the temperature of the hole wall.

21. The system according to any one of claims 16 to 20 or any other claim herein, wherein, The processing protocol includes setting the speed and / or direction and / or duration of rotation of the at least one main shaft.

22. The system according to any one of claims 16 to 21 or any other claim herein, wherein, The processing protocol is selected or input through a graphical user interface.

23. The system according to any one of claims 16 to 22 or any other claim herein, further comprising a sensor downstream of the at least one processor or microprocessor, the sensor relaying a feedback signal to the at least one processor or microprocessor.

24. The system according to any one of claims 16 to 23 or any other claim herein, wherein, The at least one main shaft is movable or deflectable between a first extended position and a second retracted position.

Images