Method, device and apparatus for separating cells by tilting

Abstract

A method includes: maintaining a container defining a first channel in a first orientation for a first time period such that a first axis defined by the first channel is in a non-vertical direction; and maintaining the container in a second orientation different from the first orientation for a second time period after the first time period. In some cases, the container contains a solution containing biological cells in the first channel.

Description

[0001] Related applications This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 520,923, filed August 21, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0002] This application relates to methods, apparatus, and equipment for separating biological cells or other particles. Background Technology

[0003] The separation of biological cells is a crucial step in many biological processes and assays. Conventional methods for separating biological cells involve subjecting the cells in a liquid to significant acceleration, such as through centrifugation. Centrifugal force causes the cells to travel to the bottom of the tube and form a precipitate, allowing the remaining liquid to separate from the precipitate.

[0004] However, the mechanical forces applied to biological cells and the formation of deposits can affect them. For example, excessive centrifugal force can cause biological cells to lyse. Summary of the Invention

[0005] Therefore, there is a need for methods, apparatus, and devices that do not require high acceleration to separate biological cells. Such methods, apparatus, and devices can replace conventional methods, apparatus, and devices used for separating biological cells. These methods, apparatus, and devices can improve the accuracy or efficiency of biological processes and assays by better preserving the physical and biological properties of cells during separation by reducing or eliminating the need for applying high acceleration or force. Such methods, apparatus, and devices can also be used to wash other types of samples, such as beads or particles bound to biomolecules.

[0006] Several embodiments that overcome the limitations and drawbacks of existing methods, apparatuses, and devices are presented in more detail below. These embodiments provide methods, apparatuses, and devices for isolating biological cells.

[0007] As described in more detail below, according to some embodiments, a method includes holding a container defining a first channel in a first orientation during a first time period, such that a first axis defined by the first channel is in a non-vertical direction. The container contains a solution containing biological cells in the first channel. The method further includes holding the container in a second orientation, different from the first orientation, during a second time period following the first time period.

[0008] According to some embodiments, an apparatus includes: a container holder for holding a container; and a tilting device coupled to the container holder for positioning the container holder in a first orientation at a first time, and positioning the container holder in a second orientation different from the first orientation at a second time different from the first time.

[0009] According to some embodiments, a method for separating non-cellular material from biological cells in a solution includes holding a container defining a first channel in a first orientation for a first time period, such that a first axis defined by the first channel is in a non-vertical direction. The container contains a solution in the first channel, the solution containing biological cells and non-cellular material. The method further includes: holding the container in a second orientation different from the first orientation for a second time period after the first time period; and aspirating a portion, but not all, of the solution.

[0010] According to some embodiments, a method includes: holding a container defining a first channel in a first orientation during a first time period, such that a first axis defined by the first channel is in a first non-vertical direction, the container containing a solution containing biological cells in the first channel; and holding the container in a second orientation different from the first orientation during a second time period after the first time period, such that the first axis defined by the first channel is in a second non-vertical direction different from the first non-vertical direction. Attached Figure Description

[0011] To better understand the foregoing embodiments and additional embodiments, the following description of the embodiments should be referred to in conjunction with the following drawings, in which the same element symbols are used throughout the drawings to refer to corresponding parts.

[0012] Figure 1 This is a perspective view of a plate according to some embodiments.

[0013] Figure 2 for Figure 1 The image shows a cross-sectional view of the plate.

[0014] Figure 3 A tube is illustrated according to some embodiments.

[0015] Figures 4A to 4I An apparatus for separating biological cells and its operation are illustrated according to some embodiments.

[0016] Figure 5 and 6 The experimental results obtained by using the methods described in this paper are illustrated.

[0017] Figures 7A to 7C A flowchart illustrating a method for condensing biological cells according to some embodiments is provided.

[0018] Figure 8 A flowchart illustrating a method for separating non-cellular material from biological cells according to some embodiments is provided.

[0019] Figure 9A flowchart illustrating a method for concentrating biological cells in a container defining channels, according to some embodiments.

[0020] Figure 10 A block diagram illustrating the electrical components of a device according to some embodiments.

[0021] Figure 11A and 11B A schematic diagram illustrating an impact device according to some embodiments.

[0022] The same component symbols are used throughout the diagram to refer to the corresponding parts.

[0023] Unless otherwise stated, the diagrams may not be drawn to scale. Detailed Implementation

[0024] Methods, apparatus, and devices for washing samples are described. Reference will be made to certain embodiments, examples of which are illustrated in the accompanying drawings. While the claims will be described in conjunction with embodiments, it should be understood that the claims are not intended to be limited to these particular embodiments. Rather, the embodiments are intended to cover alternatives, modifications, and equivalents within the spirit and scope of the appended claims.

[0025] Furthermore, numerous specific details are set forth in the following description to provide a thorough understanding of the embodiments. However, it will be apparent to those skilled in the art that the embodiments may be practiced without these specific details. In other instances, methods, procedures, components, and networks well known to those skilled in the art have not been described in detail to avoid obscuring aspects of the embodiments.

[0026] It should be understood that although the terms first, second, etc., may be used herein to describe various elements, these elements should not be limited by these terms. These terms are only used to distinguish one element from another. For example, without departing from the scope of the embodiments, a first channel may be referred to as a second channel, and similarly, a second channel may be referred to as a first channel. Both the first channel and the second channel are channels, but they are not the same channel. Similarly, without departing from the scope of the embodiments, a first time period may be referred to as a second time period, and similarly, a second time period may be referred to as a first time period. Both the first time period and the second time period are time periods, but they are not the same time period.

[0027] The terminology used in the description of the embodiments herein is for the purpose of describing particular embodiments only and is not intended to limit the invention. As used in the description of the embodiments and the appended claims, the singular forms “a,” “an,” and “the” are also intended to include the plural forms unless the context clearly indicates otherwise. It should also be understood that the term “and / or,” as used herein, refers to and covers any and all possible combinations of one or more of the associated listed items. It will be further understood that, when used in this specification, the terms “comprises” and / or “comprising” specify the presence of the stated features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.

[0028] Figure 1 This is a perspective view of a container (e.g., plate 100) according to some embodiments. Plate 100 has a top surface 120 and a bottom surface 130 opposite to the top surface 120. A plurality of holes 112 (e.g., holes 112-1 to holes 112-8) are defined in plate 100. Plate 100 includes a first portion 140 corresponding to the bottom of the plurality of holes 112 and a second portion 150 corresponding to one or more walls of the plurality of holes 112. In some embodiments, plate 100 is integrally formed. In some embodiments, plate 100 is formed by attaching two or more portions together (e.g., by joining separately formed first portions 140 and second portions 150). In some embodiments, as Figure 1 As shown, multiple holes 112 are arranged in an array (e.g., a 2×3 array, a 2×4 array, a 3×4 array, a 4×6 array, a 6×8 array, an 8×12 array, a 16×24 array, a 32×48 array, etc.). In some embodiments, the corresponding holes 112 are cylindrical holes (e.g., the corresponding holes 112 have a circular shape along a cross-section that is generally parallel to the plane of the plate 100). Figure 1 Line II-II is also on display. Figure 2 The cross-sectional view is taken from line II-II.

[0029] Figure 2 for Figure 1The cross-sectional view of plate 100 shown in the figure. In some embodiments, the corresponding hole 112 has a width (W) of 2 mm to 170 mm (e.g., 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, 40 mm, 45 mm, 50 mm, 55 mm, 60 mm, 65 mm, 70 mm, 75 mm, 80 mm, 85 mm, 90 mm, 95 mm, 100 mm, 110 mm, 120 mm, 130 mm, 140 mm, 150 mm, 160 mm, 170 mm or any interval between two of the foregoing values, such as 5 mm to 8 mm). In some embodiments, the corresponding hole 112 has a height (H) of 2 mm to 170 mm (e.g., 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, 40 mm, 45 mm, 50 mm, 55 mm, 60 mm, 65 mm, 70 mm, 75 mm, 80 mm, 85 mm, 90 mm, 95 mm, 100 mm, 110 mm, 120 mm, 130 mm, 140 mm, 150 mm, 160 mm, 170 mm or any interval between two of the foregoing values, such as 3 mm to 70 mm).

[0030] Figure 2 Also shown are a first axis 202 defined by a first channel (e.g., hole 112-1) and a second axis 204 defined by a second channel (e.g., hole 112-2). In some embodiments, the first axis 202 is parallel to the longitudinal direction of the first channel (e.g., hole 112-1). In some embodiments, the first axis 202 passes through the center of the first channel. In some embodiments, the second axis 204 is parallel to the longitudinal direction of the second channel (e.g., hole 112-2). In some embodiments, the second axis 204 passes through the center of the second channel. In some embodiments, the first axis 202 is parallel to the height direction of the first channel (e.g., hole 112-1). In some embodiments, the second axis 204 is parallel to the height direction of the second channel (e.g., hole 112-2). In some embodiments, the first axis 202 is parallel to the second axis 204. In some embodiments, the first axis 202 is not parallel to the second axis 204 (e.g., the first axis 202 and the second axis 204 form a predefined non-zero angle, such as 1 degree, 5 degrees, 10 degrees, 15 degrees, 20 degrees or between any two angles selected from the aforementioned angles).

[0031] although Figure 1 and 2 The illustration depicts a plate with multiple holes, but alternatively or additionally, a container with only a single hole or channel (e.g., a pipe) may be used.

[0032] Figure 3 A pipe 300 is illustrated according to some embodiments. The pipe 300 includes a pipe wall 310 defining a channel 302. The channel 302 has or defines an axis 318. In some embodiments, the axis 318 is parallel to the longitudinal direction of the channel 302.

[0033] Figure 3 The illustration also shows that tube 300 in some cases contains a solution 320 containing biological cells 330 or particles. However, as those skilled in the art will recognize, solution 320 and biological cells 330 are not part of tube 300.

[0034] Figures 4A to 4H The illustration depicts an apparatus 400 for separating biological cells and an operation for separating biological cells according to some embodiments.

[0035] exist Figure 4A In this embodiment, the device includes a container holder 410 and a tilting device 420. In some embodiments, the device further includes an impact device 430.

[0036] like Figure 4A As shown, in operation, a container 300 (e.g., a tube) containing a solution 320 having biological cells 330 or particles can be placed in a container holder 410.

[0037] In some embodiments, the container holder 410 is in a second orientation. In some embodiments, the second orientation is as follows: Figure 4A The container holder 410 is generally vertically oriented as shown in the figures. In some embodiments, the container holder 410 is considered to be generally vertically oriented when the channel defined by the container held by the container 410 is defined as an axis that is generally vertically oriented. In some embodiments, the container holder 410 (e.g., a tube holder) is considered to be generally vertically oriented when the channel defined by the container holder 410 (e.g., a channel in which a tube is inserted) is defined as an axis that is generally vertically oriented. For example, a first axis 318 defined by the channel 302 of the container 300 or the channel of the container holder 410 is generally vertical (e.g., axis 318 is generally parallel to the vertical direction 440). In some embodiments, the container holder 410 (e.g., a plate holder) is considered to be generally vertically oriented when the bottom surface of the container holder 410 is generally perpendicular to the vertical direction 440.

[0038] Figure 4A It also shows that the container holder 410 can be rotated or tilted by the tilting device 420.

[0039] Figure 4B The container holder 410 is illustrated in a first orientation. In some embodiments, the first orientation is as follows: Figure 4B The non-vertical orientation is shown in the figure. In some embodiments, the container holder 410 is considered to be in a non-vertical orientation when the channel defined by the container held by the container 410 is defined on an axis that is in a non-vertical orientation. In some embodiments, the container holder 410 (e.g., a tube holder) is considered to be in a non-vertical orientation when the channel defined by the container holder 410 (e.g., a channel in which a tube is inserted) is defined on an axis that is in a non-vertical orientation (e.g., the axis has a non-zero angle relative to the vertical direction). For example, a first axis 318 defined by the channel 302 of the container 300 or the channel of the container holder 410 is non-vertical (e.g., axis 318 is not parallel to the vertical direction 440). In some embodiments, the container holder 410 (e.g., a plate holder) is considered to be in a non-vertical orientation when the bottom surface of the container holder 410 is not perpendicular to the vertical direction 440.

[0040] Figure 4C The container holder 410 is held in a first orientation. The inclination of the channel defined by the container 310 accelerates the settling of biological cells or particles.

[0041] Figure 4D The demonstration continues to hold the container holder 410 in the first orientation, such that the tilt of the channel defined by the container 310 continues to promote the sedimentation of biological cells or particles.

[0042] Figure 4D It also demonstrates that the container holder 410 can be rotated or tilted backward by the tilting device 420.

[0043] Figure 4E The container holder 410 is shown in the second orientation. Figure 4E In this process, the biological cells 330 or particles settle downwards to the bottom of the container. In some cases, because the biological cells 330 or particles settle when the container is tilted, they may have settled asymmetrically (e.g., when the container is positioned vertically, the biological cells 330 or particles may be more stacked at one end than at the opposite end). For example, when the container holder 410 or container 300 is in a tilted orientation, Figure 4E When the vertical orientation is as shown, the top surface defined by the settled biological cells 330 or particles may not be perpendicular to the vertical direction.

[0044] Figure 4FIn some embodiments, one or more mechanical impacts are illustrated to the container 310 or container holder 410. In some cases, the one or more mechanical impacts cause the biological cells 330 or particles to redistribute. In some cases, the redistribution of the biological cells 330 or particles causes the container holder 410 or container 300 to be subjected to, for example, Figure 4F When the vertical orientation is as shown, the top surface defined by the settled biological cells 330 or particles is generally perpendicular to the vertical direction.

[0045] Figure 4G Aspirator 450 or its tip is illustrated. In some embodiments, aspirator 450 aspirates at least a portion of solution 320 (and any substance that does not settle downwards with biological cells 330 or particles, such as non-cellular material).

[0046] Figure 4H Draw a portion (e.g., the majority) of the aspirated solution 320. Leave the remaining portion 330-2 of the solution 320, containing biological cells 330, in container 300. Still... Figure 4H The illustration depicts the aspirated portion 320-1 to show that the aspirated portion 320-1 has no biological cells or has a low concentration of biological cells, while the remaining portion 330-2 has a high concentration of biological cells.

[0047] Figure 4I The applicator 460 or its tip is illustrated. In some embodiments, the applicator 460 is different from the aspirator 450. In some embodiments, the applicator 460 dispenses a different solution (e.g., a wash buffer), which is mixed with the remainder of solution 320 to form a mixed solution 470. By providing a different solution, the concentration of any substance (such as non-cellular material) that did not settle downwards with the biological cells 330 or particles is reduced.

[0048] In some embodiments, the aspiration of a portion of the solution in the container and the application of additional solution are repeated to further reduce the concentration of any substances (such as non-cellular substances) that have not settled downwards with the biological cells 330 or particles.

[0049] In some embodiments, applicator 460 or another applicator provides one or more reagents. For example, in some embodiments, applicator 460 or another applicator provides a lysis buffer. In some cases, providing a lysis buffer causes at least some biological cells (e.g., red blood cells) in a solution (e.g., whole blood) to lyse. In some embodiments, a lysis buffer is provided at any stage described herein (e.g., before tilting).

[0050] Although drawn with tubes Figures 4A to 4I The operation can be performed within the system, but a board (e.g.,) can be used. Figure 1 and2 The corresponding operation is performed using plate 100 shown in the image. For the sake of brevity, such details will not be repeated here. Using plates with multiple channels allows for the simultaneous separation of multiple samples or solutions.

[0051] Example of a whole blood washing procedure Add a whole blood sample to a container (e.g., a well plate) and add lysis buffer.

[0052] Tilt the container and hold it in the tilted orientation for 35 minutes to allow particles in the whole blood and lysis buffer mixture to settle.

[0053] After 35 minutes, tap one side of the board (e.g., using a tapping block without a hammer).

[0054] The container is returned to an upright orientation (e.g., a horizontal orientation in the case of an orifice plate, so that the channels of the orifice plate are upright) and held in an upright orientation for 10 minutes, allowing the particles in the mixture to settle further.

[0055] Draw a solution from a container (e.g., from one or more holes).

[0056] Experimental results Figure 5 and 6 The experimental results obtained by using the methods described in this paper are illustrated.

[0057] Figure 5 Cell population frequency and staining index were plotted. Whole blood was stained with TBNK reagent (BD Biosciences). The whole blood sample was split into two separate samples. One sample was washed using centrifugation (e.g., by centrifugation to form sediment, followed by aspiration of the solution and addition of fresh buffer). The other sample was washed using the method described herein without centrifugation. Results showed that cell population frequency and staining index were comparable between centrifugation-based washing and washing without centrifugation.

[0058] Figure 6 A dot plot is shown, obtained by flow cytometry analysis of cells in whole blood stained with TBNK. The dot plot shows that the centrifugation-free washing method described in this application provides clear differentiation between neutrophil and natural killer (NK) cell populations.

[0059] Figures 7A to 7C A flowchart illustrating a method 700 for condensing biological cells according to some embodiments is provided.

[0060] Method 700 includes (720) maintaining a container (e.g., pipe 300 or plate 100) defining a first channel (e.g., channel 302) in a first orientation (e.g., as shown in the image) for a first time period. Figure 4CIn the non-vertical orientation shown, such that the first axis defined by the first channel (e.g., axis 318) is in the non-vertical direction (e.g., Figure 4C (Axis 318 in the middle). The container contains a solution (e.g., solution 320) containing biological cells (e.g., cell 330) in the first channel.

[0061] In some embodiments, the method includes, (702) rotating the container to position it in the first orientation before holding it in that orientation (e.g., container 310 from...). Figure 4A The vertical orientation rotation shown in the figure Figure 4B (The non-vertical orientation shown in the image).

[0062] In some embodiments, the container is positioned (704) in a second orientation (e.g., before rotating the container to place it on a first orientation). Figure 4A )superior.

[0063] In some embodiments, the container includes (706) a tube (e.g., tube 300).

[0064] In some embodiments, the container defines (708) a plurality of different and separate channels (e.g., a plate 100 having a plurality of holes, each hole corresponding to a corresponding channel).

[0065] In some embodiments, the container includes (710) an array board (e.g., board 100).

[0066] In some embodiments, when the container is in a second orientation (e.g., Figure 4D When the first axis defined by the first channel is in the generally vertical direction (722), the first axis is in the generally vertical direction.

[0067] In some embodiments, the first time period is less than (724) one hour.

[0068] In some embodiments, the first time period is less than (726) 30 minutes.

[0069] In some embodiments, the first time period is less than (728) 15 minutes.

[0070] In some embodiments, the solution has a volume of (730) at least 50 µL. In some embodiments, the solution has a volume of (732) at least 500 µL.

[0071] In some embodiments, the first axis forms an angle of (734) at least 30 degrees with the vertical direction. In some embodiments, the first axis forms an angle of (736) approximately 45 degrees with the vertical direction. In some embodiments, the first axis forms an angle of (738) at least 60 degrees with the vertical direction.

[0072] In some embodiments, the solution contains (740) whole blood.

[0073] In some embodiments, the method includes (742) adding one or more antibodies to the solution.

[0074] In some embodiments, the method includes adding (744) lysis buffer to the solution. In some embodiments, whole blood is incubated with one or more antibodies before the addition of lysis buffer (746). In some embodiments, whole blood is incubated with one or more antibodies after the addition of lysis buffer (748).

[0075] In some embodiments, the method includes (750) rotating the container in a first orientation to position the container in a second orientation (e.g., so that in...). Figure 4C The container 310 shown in the image rotates to a non-vertical orientation. Figure 4D (Vertical orientation shown in the image).

[0076] The method further includes (760) maintaining the container in a second orientation different from the first orientation during a second time period following the first time period (e.g., as...). Figure 4D (as shown in the vertical orientation).

[0077] In some embodiments, the second time period is less than 30 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 3 minutes, 2 minutes, 1 minute, or within the interval between any two of the aforementioned values. In some embodiments, the second time period is at least 60 seconds, 50 seconds, 40 seconds, 30 seconds, 20 seconds, 10 seconds, 5 seconds, 3 seconds, 2 seconds, 1 second, or within the interval between any two of the aforementioned values. In some embodiments, the second time period is at least one second.

[0078] In some embodiments, the method includes (762) providing a mechanical impact (e.g., a tap) to the container. In some embodiments, the mechanical impact is provided directly to the container. In some embodiments, the mechanical impact is provided indirectly to the container (e.g., by providing a mechanical impact to a container holder).

[0079] In some embodiments, the method includes (764) providing a series of mechanical impacts to the container over time. For example, a sequence of multiple mechanical impacts may be provided over time.

[0080] In some embodiments, when the container is in a second orientation (e.g., Figure 4D When ), a mechanical impact (766) is applied to the container.

[0081] In some embodiments, the method includes, after holding the container in a second orientation, (770) aspirating a portion, rather than all, of the solution (e.g., Figure 4F).

[0082] In some embodiments, the aspirated portion of the solution (e.g., aspirated portion 320-1) has a first concentration of biological cells (772); and the remaining portion of the solution (e.g., remaining portion 320-2) has a second concentration of biological cells greater than the first concentration. For example, the aspirated portion has a low concentration of biological cells, and the remaining portion of the solution has a high concentration of biological cells, such as... Figure 4G shown in.

[0083] In some embodiments, Figure 8 and 9 Some of the operations described can be combined with method 700. For the sake of brevity, such details will not be repeated in this article.

[0084] Figure 8 A flowchart illustrating a method 800 for separating non-cellular material from biological cells according to some embodiments is provided.

[0085] A method 800 for separating noncellular substances from biological cells in solution includes (802) holding a container defining a first channel in a first orientation for a first time period, such that a first axis defined by the first channel is in a non-vertical direction. The container contains a solution in the first channel, the solution containing biological cells and noncellular substances.

[0086] The method further includes (804) maintaining the container in a second orientation different from the first orientation during a second time period following the first time period.

[0087] The method further includes (806) aspirating a portion, but not all, of the solution.

[0088] In some embodiments, the aspirated portion has (808) a first concentration of biological cells; and the remaining portion of the solution has a second concentration of biological cells greater than the first concentration.

[0089] In some embodiments, the aspirated portion has a third concentration of noncellular material (810); and the remaining portion of the solution has a fourth concentration of noncellular material less than the third concentration.

[0090] In some embodiments, the aspirated portion contains (812) noncellular material, and the remainder of the solution contains biological cells.

[0091] In some embodiments, Figures 7A to 7C Some of the operations described in 9 can be combined with method 800. For the sake of brevity, such details will not be repeated in this article.

[0092] Figure 9A flowchart illustrating a method 900 for concentrating biological cells in a container defining channels, according to some embodiments.

[0093] Method 900 includes (902) holding a container defining a first channel in a first orientation for a first time period, such that a first axis defined by the first channel is in a first non-vertical direction. The container contains a solution containing biological cells in the first channel.

[0094] The method further includes (904) maintaining the container in a second orientation different from the first orientation during a second time period following the first time period, such that the first axis defined by the first channel is in a second non-vertical direction different from the first non-vertical direction.

[0095] In some embodiments, the first axis in the first non-vertical direction forms a first angle (906) with the vertical direction; and the first axis in the second non-vertical direction forms a second angle with the vertical direction, the second angle being smaller than the first angle.

[0096] In some embodiments, Figures 7A to 7C Some of the operations described in 8 can be combined with method 900. For the sake of brevity, such details will not be repeated in this article.

[0097] Figure 10 A block diagram illustrating the electrical components of a device according to some embodiments.

[0098] The device includes one or more processors 1002 (central processing unit, application processing unit, application-specific integrated circuit, etc.), said one or more processors and storage for performing any of the methods described herein (e.g., regarding...). Figures 7A to 7C The computer-readable storage medium 1012 (e.g., a transient computer-readable storage medium or a non-transient computer memory device, such as random access memory, read-only memory, static random access memory, and other non-volatile memory, as well as other storage devices, such as hard disk drives, optical disks, magnetic tape recording, or any combination thereof) communicates (e.g., via one or more communication buses 1008 interconnecting multiple electronic components of the device) with instructions for the operations described in 8 and 9. For example, in some embodiments, the computer-readable storage medium 1012 stores the following programs, modules, instructions, and data structures or subsets thereof: Operating system 1014 contains programs for handling various basic system services and for performing hardware-dependent tasks; A communication module (or instruction) 1016 is used to connect the controller 1006 to other electronic devices via one or more communication interfaces 1004 and one or more communication networks (e.g., wired and / or wireless communication networks); and Actuator control instruction 1018 causes an electrical signal to be provided to one or more actuators (directly from one or more processors 1002 or via driver 1010).

[0099] In some embodiments, the device includes one or more communication interfaces 1004 for communicating with other electronic devices.

[0100] In some embodiments, the controller 1006 includes one or more drivers 1010 or is electrically coupled to one or more drivers 1010 (via a system bus or any suitable circuitry). In some embodiments, one or more drivers 1010 receive instructions and / or data from one or more processors 1002 and relay the instructions and / or electrical signals to one or more actuators, such as a first actuator 1032 (e.g., in the tilting device 420), a second actuator 1034 (e.g., in the impact device 430), etc.

[0101] In some embodiments, the device includes or communicates with one or more user interface (UI) devices 1008. In some embodiments, the UI device 1008 includes one or more user input devices (e.g., keyboard, mouse, touch-sensitive surface, button, switch, etc.) for receiving user input and / or one or more output devices (e.g., display, one or more indicators, audio devices, etc.) for providing output to the user (e.g., the status of device operation).

[0102] although Figure 10 The demonstration shows the presence of a processor 1002, but in some embodiments, the device includes an additional processor. Similarly, although... Figure 10 This illustration shows one unit of computer-readable storage medium 1012, but in some embodiments, the computer-readable storage medium 1012 is implemented across multiple physical devices. In some embodiments, one or more processors 1002 communicate with one or more user interface devices (e.g., displays and one or more user input devices, such as keyboards, mice, touchscreens, etc.) for presenting information and / or receiving user input. Although Figure 10 One or more communication interfaces 1004, controllers 1006, and drivers 1010 are illustrated as separate devices, but in some embodiments, at least some of the communication interfaces 1004, controllers 1006, and drivers 1010 may be combined or integrated.

[0103] Figure 11A and 11B A schematic diagram illustrating an impact device according to some embodiments.

[0104] Figure 11A The diagram illustrates an impact device including hammer 1102. Figure 11A In this embodiment, the impact device includes an actuator 1104 coupled to a hammer 1102. The actuator 1104 moves the hammer 1102 such that the hammer 1102 can provide a mechanical impact (e.g., by raising and releasing the hammer 1102 to cause it to fall and provide a mechanical impact, or by moving the hammer 1102 forward to provide a mechanical impact). In some embodiments, the impact device includes a striking block (e.g., a block without a hammerhead) instead of the hammer 1102 for providing a mechanical impact.

[0105] Figure 11B An impact device comprising an electromagnetic coil 1112 and an actuator 1114 is illustrated. In some embodiments, the actuator 1114 comprises a magnet. The electromagnetic coil 1112 is electrically coupled to a power supply 1118 (e.g., a driver 1010, a controller 1006, etc.). Based on an electrical signal from the power supply 1118, the electromagnetic coil 1112 moves the actuator 1114 to cause the impact device to deliver a mechanical impact.

[0106] In light of these principles and examples, we now turn to certain implementation methods.

[0107] According to some embodiments, the device (e.g., device 400) includes a container holder (e.g., container holder 410) for holding a container and a tilting device (e.g., tilting device 420) coupled to the container holder for positioning the container holder in a first orientation at a first time and positioning the container holder in a second orientation different from the first orientation at a second time different from the first time.

[0108] In some embodiments, the device includes: one or more processors (e.g., processor 1002) communicating with a tilting device (e.g., actuator 1032 in the tilting device); and a memory (e.g., computer-readable storage medium 1012) storing instructions for execution by the one or more processors. The stored instructions include instructions for: sending a first set of one or more signals to the tilting device to position a container holder in a first orientation; and sending a second set of one or more signals to the tilting device to position the container holder in a second orientation.

[0109] In some embodiments, the stored instructions include instructions for: sending a first set of one or more signals to the tilting device to place the container holder in a first orientation at a first time; and sending a second set of one or more signals to the tilting device to place the container holder in a second orientation at a second time after the first time, such that the container holder remains in the first orientation for a first time period.

[0110] In some embodiments, the first time period is less than one hour.

[0111] In some embodiments, the first time period is less than 30 minutes.

[0112] In some embodiments, the first time period is less than 15 minutes.

[0113] In some embodiments, after sending the second set of one or more signals, the device will not send one or more sets of signals for positioning the container holder in any orientation other than the second orientation to the tilting device for at least a second time period from the time the second set of one or more signals were sent.

[0114] In some embodiments, the stored instructions include instructions for: sending one or more signals to the tilting device to place the container holder in a second orientation before sending a first set of one or more signals to the tilting device to place the container holder in a first orientation.

[0115] In some embodiments, the container holder includes a tube rack (e.g., container holder 410).

[0116] In some embodiments, the container includes a tube (e.g., tube 300).

[0117] In some embodiments, the container holder includes a holder for the array panel.

[0118] In some embodiments, the container includes an array board.

[0119] In some embodiments, the device includes a suction device (e.g., suction device 450) for aspirating a portion of a solution defined in a first channel within a container.

[0120] In some embodiments, the device includes an applicator (e.g., applicator 460) for applicating a washing buffer into a solution defined in a first channel in the container.

[0121] In some embodiments, the device includes a liquid transferor (e.g., a pipette). The liquid transferor can both aspirate and dispense liquids.

[0122] In some embodiments, the device includes one or more mechanical or electromechanical components (e.g., impact device 430 or one or more components thereof) for providing mechanical impact to the container.

[0123] In some embodiments, one or more mechanical or electromechanical components include a hammer (e.g., hammer 1102).

[0124] In some embodiments, one or more mechanical or electromechanical components include an electromagnetic coil (e.g., coil 1112) and an actuator (e.g., actuator 1114).

[0125] In some embodiments, the container defines a first channel along a first axis. When the container holder is in a first orientation, the container is in the first orientation; and the first axis is in a non-vertical direction.

[0126] In some embodiments, when the container holder is in the second orientation, the container is in the second orientation; and the first axis is in a generally vertical direction.

[0127] In some embodiments, when the container holder is in a first orientation, the first axis is at least 30 degrees to the vertical direction.

[0128] In some embodiments, the first axis is at least 30 degrees to the vertical direction.

[0129] In some embodiments, the first axis is at approximately 45 degrees to the vertical direction.

[0130] In some embodiments, the first axis is at least 60 degrees to the vertical direction.

[0131] In some embodiments, the device includes an applicator for dispensing one or more antibodies.

[0132] In some embodiments, the device includes an applicator for applying lysis buffer.

[0133] For purposes of explanation, the foregoing description has been described with reference to specific embodiments. However, the above illustrative discussion is not intended to be exhaustive or to limit the embodiments to the precise forms disclosed. In view of the foregoing teachings, many modifications and variations are possible. The embodiments were chosen and described in order to best explain the principles of the invention and its practical application, thereby enabling those skilled in the art to best utilize the invention and its various embodiments with various modifications suitable for the particular intended use.

[0134] Some embodiments are described with reference to the following items.

[0135] Clause 1. A method comprising: During a first time period, the container defining the first channel is held in a first orientation such that the first axis defined by the first channel is in a non-vertical direction, and the container contains a solution containing biological cells in the first channel; and During the second time period following the first time period, the container is kept in a second orientation, which is different from the first orientation.

[0136] Clause 2. The method according to Clause 1, wherein when the container is in the second orientation, the first axis defined by the first channel is generally vertical.

[0137] Article 3. The method according to Article 1 or 2 further includes: Rotate the container in the first orientation to position it in the second orientation.

[0138] Article 4. The method according to any one of Articles 1 to 3, further comprising: Rotate the container to position it in the first orientation before holding it in that orientation.

[0139] Article 5. The method according to Article 4, wherein: The container is positioned in a second orientation before being rotated to place it in the first orientation.

[0140] Article 6. The method according to any one of Articles 1 to 5, wherein: The first time period is less than one hour.

[0141] Article 7. The method according to Article 6, wherein: The first time period is less than 30 minutes.

[0142] Article 8. The method according to Article 7, wherein: The first time period is less than 15 minutes.

[0143] Article 9. The method according to any one of Articles 1 to 8, wherein: The solution has a volume of at least 50 µL.

[0144] Article 10. The method according to Article 9, wherein: The solution has a volume of at least 500 µL.

[0145] Article 11. The method according to any one of Articles 1 to 10, wherein: The first axis is at least 30 degrees to the vertical direction.

[0146] Article 12. The method according to any one of Articles 1 to 11, wherein: The first axis is approximately 45 degrees to the vertical direction.

[0147] Article 13. The method according to any one of Articles 1 to 11, wherein: The first axis is at least 60 degrees to the vertical direction.

[0148] Article 14. The method according to any one of Articles 1 to 13, wherein: The solution contains whole blood.

[0149] Article 15. The method according to Article 14 further comprises: Add one or more antibodies to the solution.

[0150] Article 16. The method according to Article 14 or 15 further comprises: Add lysis buffer to the solution.

[0151] Article 17. The method according to Article 16, wherein: Whole blood is incubated with one or more added antibodies before adding lysis buffer.

[0152] Article 18. The method according to Article 16, wherein: After adding lysis buffer, whole blood is incubated with one or more added antibodies.

[0153] Article 19. The method according to any one of Articles 1 to 18, further comprising: After holding the container in the second orientation, aspirate a portion of the solution, not all of it.

[0154] Article 20. The method according to Article 19, wherein: The aspirated portion of the solution contains a first concentration of biological cells; and The remaining portion of the solution contains biological cells at a second concentration greater than the first concentration.

[0155] Article 21. The method according to any one of Articles 1 to 19, wherein: The container contains tubes.

[0156] Article 22. The method according to any one of Articles 1 to 19, wherein: The container defines multiple distinct and separate channels.

[0157] Article 23. The method according to any one of Articles 1 to 20 and 22, wherein: The container contains the array board.

[0158] Article 24. The method according to any one of Articles 1 to 23, further comprising: Apply mechanical impact to the container.

[0159] Article 25. The method according to Article 24 further comprises: A series of mechanical impacts are applied to the container over time.

[0160] Article 26. The method according to Article 24 or 25, wherein: When the container is in the second orientation, a mechanical impact is applied to the container.

[0161] Article 27. An apparatus comprising: Container holders, used to hold containers; and A tilting device coupled to a container holder for positioning the container holder in a first orientation at a first time, and positioning the container holder in a second orientation different from the first orientation at a second time different from the first time.

[0162] Clause 28. The apparatus according to Clause 27, further comprising: One or more processors that communicate with the tilting device; and Memory that stores instructions for execution by one or more processors, including instructions for the following operations: Sending one or more signals to the tilting device to position the container holder in a first orientation; and One or more signals from a second set are sent to the tilting device to position the container holder in a second orientation.

[0163] Clause 29. The apparatus according to Clause 28, wherein: The stored instructions include instructions for the following operations: Sending one or more signals from a first group to the tilting device to position the container holder in a first orientation at a first time; and One or more signals from a second set are sent to the tilting device to place the container holder in a second orientation at a second time after the first time, such that the container holder remains in the first orientation during the first time period.

[0164] Article 30. The apparatus according to Article 29, wherein: The first time period is less than one hour.

[0165] Clause 31. The apparatus according to Clause 30, wherein: The first time period is less than 30 minutes.

[0166] Clause 32. The apparatus according to Clause 31, wherein: The first time period is less than 15 minutes.

[0167] Article 33. The equipment according to any one of Articles 28 to 32, wherein: After sending the second set of one or more signals, the device will not send one or more sets of signals for positioning the container holder in any orientation other than the second orientation to the tilting device for at least a second time period from the date of sending the second set of one or more signals.

[0168] Article 34. The equipment according to any one of Articles 28 to 33, wherein: The stored instructions include instructions for the following operations: Before sending one or more signals to the tilting device to position the container holder in a first orientation, one or more signals are sent to the tilting device to position the container holder in a second orientation.

[0169] Article 35. The equipment according to any one of Articles 27 to 34, wherein: The container holder includes a tube rack.

[0170] Article 36. The equipment according to any one of Articles 27 to 35, wherein: The container contains tubes.

[0171] Article 37. The equipment according to any one of Articles 27 to 36, wherein: The container holder includes a holder for the array plate.

[0172] Article 38. The equipment according to any one of Articles 27 to 37, wherein: The container contains the array board.

[0173] Article 39. The equipment according to any one of Articles 27 to 38, further comprising: A suction device, used to aspirate a portion of a solution defined in a first channel within a container.

[0174] Clause 40. The equipment according to any one of Clauses 27 to 39, further comprising: A dispenser for dispensing washing buffer into a solution defined in a first channel within a container.

[0175] Clause 41. The equipment according to any one of Clauses 27 to 40, further comprising: One or more mechanical or electromechanical components used to deliver a mechanical impact to a container.

[0176] Clause 42. The apparatus according to Clause 41, wherein: One or more mechanical or electromechanical components contain a hammer.

[0177] Clause 43. The apparatus according to Clause 41 or 42, wherein: One or more mechanical or electromechanical components include an electromagnetic coil and an actuator.

[0178] Article 44. The equipment according to any one of Articles 27 to 43, wherein: The container defines the first channel along the first axis; When the container holder is in the first orientation: The container is in the first orientation; and The first axis is in a non-vertical direction.

[0179] Clause 45. The apparatus according to Clause 44, wherein: When the container holder is in the second orientation: The container is in the second orientation; and The first axis is in a generally vertical direction.

[0180] Clause 46. The apparatus according to Clause 44 or 45, wherein: When the container holder is in the first orientation, the first axis is at least 30 degrees to the vertical direction.

[0181] Clause 47. The apparatus according to Clause 46, wherein: The first axis is at least 30 degrees to the vertical direction.

[0182] Clause 48. The apparatus according to Clause 47, wherein: The first axis is approximately 45 degrees to the vertical direction.

[0183] Clause 49. The apparatus according to Clause 47, wherein: The first axis is at least 60 degrees to the vertical direction.

[0184] Clause 50. The equipment according to any one of Clauses 27 to 49, further comprising: An applicator, used to apply one or more antibodies.

[0185] Clause 51. The equipment according to any one of Clauses 27 to 50, further comprising: Applicator, used to apply lysis buffer.

[0186] Article 52. A method for separating non-cellular substances from biological cells in solution, the method comprising: During a first time period, the container defining the first channel is held in a first orientation such that the first axis defined by the first channel is in a non-vertical direction, and the container contains a solution in the first channel, the solution containing biological cells and said non-cellular substances; During the second time period following the first time period, the container will be held in a second orientation different from the first orientation; and Aspirate a portion of the solution, not all of it.

[0187] Article 53. The method according to Article 52, wherein: The aspirated portion contains a first concentration of biological cells; and The remaining portion of the solution contains biological cells at a second concentration greater than the first concentration.

[0188] Article 54. The method according to Article 52 or 53, wherein: The aspirated portion contained a third concentration of non-cellular material; and The remaining portion of the solution contains a fourth concentration of non-cellular substances, which is less than the third concentration.

[0189] Article 55. The method according to any one of Articles 52 to 54, wherein: The aspirated portion contains non-cellular material, while the remainder of the solution contains biological cells.

[0190] Article 56. A method comprising: During a first time period, the container defining the first channel is held in a first orientation, such that the first axis defined by the first channel is in a first non-vertical direction, and the container contains a solution containing biological cells in the first channel; and During a second time period following the first time period, the container is held in a second orientation different from the first orientation, such that the first axis defined by the first channel is in a second non-vertical direction different from the first non-vertical direction.

[0191] Article 57. The method according to Article 56, wherein: The first axis in the first non-vertical direction forms a first angle with the vertical direction; and The first axis in the second non-vertical direction forms a second angle with the vertical direction, and the second angle is smaller than the first angle.

Claims

1. A method comprising: During a first time period, the container defining the first channel is held in a first orientation such that the first axis defined by the first channel is in a non-vertical direction, and the container contains a solution containing biological cells in the first channel; and During a second time period following the first time period, the container is held in a second orientation, different from the first orientation.

2. The method of claim 1, wherein when the container is in the second orientation, the first axis defined by the first channel is in a generally vertical direction.

3. The method according to claim 1, further comprising: The container is rotated in the first orientation to position it in the second orientation.

4. The method of claim 1, further comprising: Before holding the container in the first orientation, rotate the container to position it in the first orientation.

5. The method according to claim 4, wherein: The container is positioned in the second orientation before being rotated to place it in the first orientation.

6. The method according to claim 1, wherein: The solution contains whole blood.

7. The method of claim 6, further comprising: One or more antibodies are added to the solution.

8. The method of claim 6, further comprising: Add lysis buffer to the solution.

9. The method according to claim 8, wherein: After adding the lysis buffer, the whole blood is incubated with one or more added antibodies.

10. The method of claim 1, further comprising: After holding the container in the second orientation, a portion, but not all, of the solution is aspirated.

11. The method of claim 10, wherein: The aspirated portion of the solution contains a first concentration of the biological cells; and The remaining portion of the solution contains biological cells at a second concentration greater than the first concentration.

12. The method of claim 1, further comprising: A mechanical impact is applied to the container.

13. The method of claim 12, further comprising: A series of mechanical impacts are applied to the container over time.

14. The method according to claim 12, wherein: The mechanical impact is provided to the container when it is in the second orientation.

15. An apparatus comprising: Container holder, used to hold containers in place; as well as A tilting device coupled to the container holder for positioning the container holder in a first orientation at a first time, and positioning the container holder in a second orientation different from the first orientation at a second time different from the first time.

16. The device according to claim 15, further comprising: One or more processors that communicate with the tilting device; as well as A memory that stores instructions for execution by the one or more processors, the stored instructions including instructions for the following operations: Sending one or more signals in the first group to the tilting device to position the container holder in the first orientation; and A second set of one or more signals is sent to the tilting device to position the container holder in the second orientation.

17. The apparatus according to claim 16, wherein: The stored instructions include instructions for the following operations: Send one or more signals from the first group to the tilting device to place the container holder on the first orientation at a first time; as well as One or more signals from the second group are sent to the tilting device to position the container holder in the second orientation at a second time after the first time, such that the container holder remains in the first orientation during the first time period.

18. The apparatus according to claim 16, wherein: After sending the second set of one or more signals, the device will not send one or more sets of signals for positioning the container holder in any orientation other than the second orientation to the tilting device for at least a second time period from the date of sending the second set of one or more signals.

19. The apparatus according to claim 16, wherein: The stored instructions include instructions for the following operations: Before sending one or more signals from the first set to the tilting device for positioning the container holder in the first orientation, one or more signals are sent to the tilting device for positioning the container holder in the second orientation.

20. The device according to claim 16, further comprising: A suction device for aspirating a portion of a solution defined in a first channel within the container; or A dispenser for dispensing washing buffer into a solution defined in a first channel within the container.

21. The device according to claim 16, further comprising: One or more mechanical or electromechanical components for providing mechanical impact to the container.

22. The device according to claim 21, wherein: The one or more mechanical or electromechanical components include a hammer.

23. The device according to claim 21, wherein: The one or more mechanical or electromechanical components include electromagnetic coils and actuators.

24. A method for separating non-cellular substances from biological cells in solution, the method comprising: During a first time period, the container defining the first channel is held in a first orientation such that the first axis defined by the first channel is in a non-vertical direction, and the container contains the solution in the first channel, the solution containing the biological cells and the non-cellular substances; During a second time period following the first time period, the container is held in a second orientation different from the first orientation. as well as Aspirate a portion of the solution, not all of it.

25. The method of claim 24, wherein: The aspirated portion contains a first concentration of the said biological cells; and The remaining portion of the solution contains biological cells at a second concentration greater than the first concentration.

26. The method of claim 24, wherein: The aspirated portion contains a third concentration of the aforementioned non-cellular substance; and The remaining portion of the solution contains a fourth concentration of the non-cellular substance that is less than the third concentration.

27. The method of claim 24, wherein: The aspirated portion contains the non-cellular material, and the remainder of the solution contains the biological cells.

28. A method comprising: During a first time period, the container defining the first channel is held in a first orientation such that a first axis defined by the first channel is in a first non-vertical direction, and the container contains a solution containing biological cells in the first channel; and During a second time period following the first time period, the container is held in a second orientation different from the first orientation, such that the first axis defined by the first channel is in a second non-vertical direction different from the first non-vertical direction.

29. The method according to claim 28, wherein: The first axis in the first non-vertical direction forms a first angle with the vertical direction; and The first axis in the second non-vertical direction forms a second angle with the vertical direction, and the second angle is smaller than the first angle.

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