Cell detachment device and cell detachment method

Abstract

To provide a cell detachment device and a cell detachment method which are capable of efficiently detaching cells from a culture vessel in a non-contact manner.SOLUTION: A cell detachment device 10 detaches cells 1 from a cell adhesion surface of a culture vessel 2, and comprises: a plurality of vibrators 11 which irradiate the cells 1 with ultrasonic beams 13 via the culture vessel 2; element driving units 14 which apply voltages to piezoelectric elements 411 of the vibrators 11; a scanning mechanism 15 which relatively moves the positions of the vibrators 11 relative to the culture vessel 2; and a control unit 16 which controls the presence or absence of the output of the ultrasonic beams 13, the beam output intensity, and the positions of the vibrators 11, wherein the vibrators 11 are arranged along any arrangement axis direction, and the scanning mechanism 15 scans the vibrators 11 along a scanning axis direction different from the arrangement axis direction.SELECTED DRAWING: Figure 1A

Description

[Technical field]

[0001] The present invention relates to a cell detachment device and a cell detachment method. [Background technology]

[0002] In the application of regenerative medicine, treatments using cultured cells have been put to practical use. Cells are grown in specific cell culture vessels (hereinafter sometimes simply referred to as "culture vessels"), including cell culture dishes and plates, multi-well plates, microtiter plates, flasks, multi-cell culture flasks, and multi-layer cell culture vessels. The cells grow by adhering to the bottom surface of the culture vessel. The grown cells form aggregates such as cell sheets, which are used for treatments, etc.

[0003] As a method for detaching cells adhered to a culture vessel from the vessel, for example, the technique described in Patent Document 1 is known.

[0004] Patent Document 1 describes a cell detachment device that detaches cultured cells from a culture vessel to which the cells are attached. This cell detachment device includes an ultrasonic probe, a moving mechanism, a voltage application unit, a cell state detection unit, and a detachment planning unit. In this device, the ultrasonic probe has a number of piezoelectric elements that generate ultrasonic waves in response to an applied voltage and produce an ultrasonic beam. The movement mechanism moves the ultrasonic probe, making it possible to change the relative position between the ultrasonic probe and the culture vessel. The voltage application unit is connected to the plurality of piezoelectric elements and applies a voltage to the plurality of piezoelectric elements with a delay time. The cell state detection unit detects the state of the cells in the culture vessel. The ablation planning unit refers to the cell state obtained by the cell state detection unit and determines the transmission conditions of the ultrasonic beam.

[0005] That is, Patent Document 1 describes a technique for detaching cells by moving an ultrasonic probe relative to a cell culture vessel. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2023-069062 A Summary of the Invention [Problem to be solved by the invention]

[0007] However, the cell detachment device described in Patent Document 1 leaves room for improvement. For example, when the area of ​​a culture vessel is enlarged to culture a large amount of cells, the cell detachment device described in Patent Document 1 takes time to evenly detach the entire culture vessel, and there are concerns that this is not efficient. Therefore, it is considered desirable to improve this.

[0008] The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a cell detachment device and a cell detachment method that can efficiently detach cells from a culture vessel in a non-contact manner. [Means for solving the problem]

[0009] The cell detachment device of the present invention, which solves the above-mentioned problems, is a cell detachment device that detaches cells from a cell adhesion surface of a culture vessel, and has a plurality of transducers that irradiate ultrasonic beams to the cells through the culture vessel, an element driving unit that applies a voltage to piezoelectric elements of the transducers, a scanning mechanism that moves the position of the transducers relatively to the culture vessel, and a control unit that controls the presence or absence of output of the ultrasonic beam, the beam output intensity, and the position of the transducers, wherein the transducers are arranged along an arbitrary array axis direction, and the scanning mechanism scans the transducers along a scanning axis direction different from the array axis direction. Effect of the Invention

[0010] The present invention can provide a cell detachment device and a cell detachment method that can efficiently detach cells from a culture vessel in a non-contact manner. Problems, configurations and effects other than those described above will become apparent from the following description of the embodiments. [Brief description of the drawings]

[0011] [Figure 1A] FIG. 1 is a schematic diagram of a cell detachment device 10 according to a first embodiment. [Figure 1B] FIG. 1 is a plan view of a cell peeling device 10 according to a first embodiment. [Figure 2A] 2 is a schematic cross-sectional view of a vibrator 11. FIG. [Figure 2B] FIG. 2 is a schematic plan view of a vibrator 11. [Figure 3A] 3 is a schematic diagram showing an example of a beam shape 310 of the ultrasonic beam 13. FIG. [Figure 3B] 13 is a schematic diagram showing another example of the beam shape 310 of the ultrasonic beam 13. FIG. [Figure 4A] FIG. 13 is an explanatory diagram showing the arrangement of the direction of major axis 321 of flat beam shape 310 and the scanning axis, and shows a configuration in which the direction perpendicular to major axis 321 is the scanning axis. [Figure 4B] FIG. 13 is an explanatory diagram showing the arrangement of the direction of major axis 321 of flat beam shape 310 and the scanning axis, showing a configuration in which the direction of major axis 321 is the scanning axis. [Figure 5A] FIG. 1 is an explanatory diagram of a case where an ultrasonic beam 13 is irradiated onto the inner region of a cell sheet adhered to a cell adhesive surface on the inner side of a culture vessel 2. [Figure 5B] FIG. 13 is an explanatory diagram of a case where an ultrasonic beam 13 is irradiated onto the edge of a cell sheet. [Figure 6] FIG. 2 is a schematic diagram of the cell detachment device 10, illustrating the control unit 16 in detail. [Figure 7] 2 is an explanatory diagram illustrating an example of a voltage waveform output from an element driving unit 14 and applied to each transducer 11 constituting an ultrasonic probe 12. FIG. [Figure 8] 5 is a schematic diagram showing cells 1 on a cell culture surface in a culture vessel 2, with detached regions 511 and non-detached regions 512. FIG. [Figure 9] 3 is a schematic diagram showing an example of time-series control of the transducer 11 in the first embodiment. FIG. [Figure 10A] 2 is a schematic diagram for explaining an example of an arrangement axis and a scanning axis of a transducer 11. FIG. [Figure 10B] 13 is a schematic diagram for explaining another example of the arrangement axis and scanning axis of the transducer 11. FIG. [Figure 10C] 13 is a schematic diagram for explaining another example of the arrangement axis and scanning axis of the transducer 11. FIG. [Figure 10D] 13 is a schematic diagram for explaining another example of the arrangement axis and scanning axis of the transducer 11. FIG. [Figure 11] FIG. 2 is a schematic diagram of a cell detachment device 10 according to a first modified example of the first embodiment. [Figure 12] FIG. 2 is a schematic diagram showing a configuration example of a cell detachment device 10 according to a first modified example of the first embodiment, in which a configuration example of a control unit 16 is particularly explained in detail. [Figure 13] 10A to 10C are schematic diagrams showing an example of time-series control of the transducer 11 in the first modified example of the first embodiment. [Figure 14] FIG. 2 is a schematic diagram of a cell detachment device 10 according to a second modified example of the first embodiment. [Figure 15] FIG. 1 is a schematic diagram of a cell detachment device 10 according to a second embodiment. [Figure 16] FIG. 2 is a schematic diagram showing an example of a database 52 having ultrasonic irradiation conditions. [Figure 17] 1 is a flowchart illustrating one aspect of a cell detachment method according to an embodiment of the present invention. [Figure 18] 10 is a flowchart illustrating another aspect of the cell detachment method according to the present embodiment. [Figure 19] 1 is a flowchart illustrating the contents of a specific example of a cell detachment method according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a cell detachment device and a cell detachment method according to an embodiment of the present invention will be described in detail with reference to the accompanying drawings. In the description of each embodiment, substantially the same or similar configurations are denoted by the same reference numerals, and where a description is redundant, the description may be omitted.

[0013] The cell detachment device and method according to the present embodiment detach cells that are in an adherent state from a cell adhesion surface, which is the culture surface of a culture vessel. In this specification, the cell adhesion surface refers to a surface to which cells adhere in a culture vessel. The cell adhesion surface may be flat or curved. The cell detachment device according to the present embodiment can be used as a part of a cell culture device. In addition, in the cell detachment method according to the present embodiment, a method of forming a cell culture medium in a commonly used cell culture dish and culturing adhesive cells is described as an example, but the scope of application is not limited to this. In addition, in this embodiment, various types of culturable adhesive cells can be targeted, and are not limited to specific cells. In addition, as the culture medium used for culturing cells in this embodiment, both a serum-free medium and a medium containing serum can be adopted.

[0014] Furthermore, the culture vessel is not limited to one layer or two layers. A one-layer culture vessel is a culture vessel in which the inner wall of the culture vessel is the cell adhesion surface. A two-layer culture vessel is a culture vessel in which, for example, a sheet-like member is provided inside the culture vessel, and not only the inner wall of the culture vessel but also the surface of the sheet-like member is the cell adhesion surface. The cell detachment device and method according to this embodiment are intended for a culture vessel having a cell adhesion surface inside the culture vessel, and may be a culture vessel provided with sheets of multiple layers. In this specification, the term "culture vessel" refers to any vessel in which cells are stored, and is not limited to vessels used for cell culture. For example, the cell detachment device and method according to the present embodiment can be applied to the case where cells are detached after being cultured in a culture vessel. The cell detachment device and method according to the present embodiment can also be applied to the case where cells are transported using a vessel (culture vessel) and then removed when necessary.

[0015] In order to explain this embodiment, a system is used in which an ultrasonic beam is irradiated upward from the outside of the bottom surface of a culture vessel in a single-layer culture state in which cells immersed in a culture medium in the culture vessel are attached to the bottom surface. However, this embodiment is not limited to this system, and can be applied to a system in which ultrasonic waves are transmitted from the side or top surface of the culture vessel through the culture medium, etc.

[0016] In the cell detachment device and method according to the present embodiment, an ultrasonic probe equipped with multiple transducers is placed on the outside of the culture vessel, for example on the outside of the bottom surface, and ultrasonic beams with a controlled beam output are irradiated from the ultrasonic probe to cells adhering to the culture surface of the culture vessel. At this time, the ultrasonic probe and the outer surface of the culture vessel may be in contact with each other via an acoustic coupler for efficiently transmitting ultrasonic waves, or a matching layer made of silicone rubber, fluororubber, or the like. In addition, the ultrasonic probe (transducer) and the outer surface of the culture vessel may not be in direct contact with each other, but may be placed with a fluid interposed between them.

[0017] In the cell detachment device and method according to the present embodiment, an ultrasonic beam having a certain intensity or more is irradiated from an ultrasonic probe to impart acoustic radiation pressure to cells. In addition, acoustic streaming may occur on the path of the ultrasonic beam due to the irradiated ultrasonic beam. An upward force is applied to the cells due to the acoustic radiation pressure and / or acoustic streaming. It has been experimentally confirmed that cells can be detached from the cell adhesion surface of a culture vessel by irradiating an ultrasonic beam with an ultrasonic intensity exceeding a certain threshold (detachment threshold). Ultrasonic forces include forces due to high-speed mechanical vibrations and forces due to sound waves. In this specification, in order to distinguish between the two, high-speed mechanical vibrations will be called "vibration force" and forces due to sound waves will be called "acoustic force." Acoustic force includes the driving force of the acoustic radiation pressure and acoustic streaming described above. The cell detachment device and method according to the present embodiment detaches cells using acoustic force.

[0018] In this embodiment, the beam output of the arbitrarily controlled ultrasonic beam may be, for example, electronically controlled ultrasonic beam intensity, ultrasonic beam transmission time (duty ratio, repetition interval), etc. The ultrasonic beam refers to a band-like or rod-like region formed by ultrasonic waves transmitted from a piezoelectric element.

[0019] The intensity of the beam output of the ultrasonic beam can be controlled, for example, by the following methods: Changing the intensity of the voltage applied to the transducer of the ultrasonic probe; Changing the number of waves (number of sine waves) of the voltage waveform to be applied; In the case of an ultrasonic probe having multiple piezoelectric elements, selecting all or some of the piezoelectric elements to which voltage is applied; Note that the voltage waveform is not limited to a sine wave and may be, for example, a rectangular wave.

[0020] The transmission time of the ultrasonic beam can be controlled, for example, by the following method. The wave number or application time of the voltage applied to the piezoelectric element of the ultrasonic probe at a specific frequency is changed. Note that the specific frequency is determined by the dimensions and material of the piezoelectric element, and is arbitrarily determined according to the piezoelectric element. According to the experiments of the present inventors, the suitable frequency for detaching cells is about 150 kHz to 50 MHz. Although the control method for the intensity and transmission time of the ultrasonic beam has been described above, the control method is not limited to the above.

[0021] In this embodiment, the ultrasonic probe includes a plurality of transducers, which are arranged in an arbitrary axial direction. In this specification, this axis is called the array axis. The plurality of transducers scan along a scanning axis that is different from the array axis and is generally perpendicular to the array axis. At this time, the transducers that generate ultrasonic beams may scan simultaneously, or may be selected in sequence to scan a single one.

[0022] The cell detachment device according to this embodiment adjusts the beam intensity of the beam output of the ultrasonic beam to a beam intensity exceeding a specific threshold (detachment threshold) based on the desired detachment area and coordinate data in the culture vessel, and locally irradiates the ultrasonic beam from outside the culture vessel through the bottom surface to cut the bond between the cell and the cell adhesion surface. The desired detachment area may be the entire culture vessel or a specific part of it.

[0023] For example, by scanning a transducer corresponding to the coordinates of the array axis of a desired ablation region on a track passing through the desired ablation region and irradiating the cells with an ultrasonic beam, it is possible to efficiently ablate only a desired region. In this embodiment, it is possible to scan multiple transducers corresponding to a desired region simultaneously or sequentially, and even a large ablation region can be efficiently ablated.

[0024] Therefore, the cell detachment device according to the present embodiment can detach cells from a culture vessel in a non-contact manner, and therefore, the cell culture device including the cell detachment device according to the present embodiment can automate the cell detachment process and achieve high throughput.

[0025] (First embodiment) Next, a specific embodiment (first embodiment) of the cell peeling device according to the present invention will be described. Fig. 1A is a schematic diagram of a cell detachment device 10 according to the first embodiment. Fig. 1B is a plan view of the cell detachment device 10 according to the first embodiment. In this specification and drawings, the directions of the x, y and z axes are shown in the drawings to show the relative positions of the drawings. In the coordinate axis display, a double circle indicates an arrow pointing from the back side of the drawing to the front side, and a cross indicates an arrow pointing from the front side of the drawing to the back side. Note that the coordinate system display of the x, y and z axes indicates the direction of the axis, and the origin position is not relevant. In addition, since the coordinate system display indicates the relationship between the directions of the drawings, it goes without saying that the direction of the x-axis is arbitrary. In this specification, the x-axis is aligned with the scanning direction (scanning axis), and the xy plane is aligned parallel to the scanning plane. The z-axis is aligned perpendicular to the xy plane.

[0026] In this embodiment, for example, the following configuration is disclosed. 1A, a cell detachment device 10 according to this embodiment is a device that detaches a cell 1 during culture by transmitting ultrasonic waves to the cell 1 that is adhered to the cell adhesion surface of a culture vessel 2. The cell detachment device 10 has an ultrasonic probe 12, an element driving unit (driving circuit) 14, a scanning mechanism 15, and a control unit 16. The ultrasonic probe 12 is connected to an element driving unit 14. The element driving unit 14 and the scanning mechanism 15 are connected to a control unit 16. The cell detachment device 10 also includes an input unit 56 and a display unit 57 (both of which are shown in FIG. 6).

[0027] The ultrasonic probe 12 has a plurality of transducers 11, and generates an ultrasonic beam 13 by an applied voltage. The ultrasonic beam 13 generated by the plurality of transducers 11 is irradiated onto the cell 1 through the culture vessel 2. The element driver 14 is connected to the vibrator 11 and applies a voltage to the piezoelectric element of the vibrator 11 . 1B, the scanning mechanism 15 moves the ultrasonic probe 12, i.e., the position of the transducer 11, relative to the culture vessel 2 by moving the ultrasonic probe 12 as shown by the arrow (scanning axis) in FIG. 1B. With this configuration, the ultrasonic probe 12 is moved, so that the cell 1 can be efficiently detached.

[0028] The control unit 16 controls whether or not the ultrasonic beam 13 is output, the beam output intensity, and the position of the transducer 11 . The control unit 16 may include an input unit 56 (see FIGS. 6, 12, and 15). The input unit 56 inputs the shape of the region of the cells 1 to be detached from the cell adhesion surface of the culture vessel 2. The control unit 16 controls the element driving unit 14 and the scanning mechanism 15 using information obtained from the input unit 56. The control unit 16 controls whether or not a voltage is applied to the transducer 11, and the strength, wave number, and repetition time of the voltage to be applied. Furthermore, the control unit 16 controls the scanning mechanism 15 based on the coordinates of the desired peeling region 511 (see FIG. 8).

[0029] In the cell detachment device 10, the transducers 11 are arranged along an arbitrary array axis direction, and the scanning mechanism 15 causes the transducers 11 to scan along a scanning axis direction different from the array axis direction. In this way, the cell detachment device 10 can scan the arranged transducers 11 in a scanning axis direction different from the array axis direction (for example, a direction perpendicular to the array axis direction). This allows the cell detachment device 10 to efficiently detach the cell 1 from the cell adhesion surface of the culture vessel 2. The cell detachment device 10 also irradiates the cell 1 with an ultrasonic beam 13 through the culture vessel 2. This allows the cell detachment device 10 to detach the cell 1 from the cell adhesion surface of the culture vessel 2 in a non-contact manner. Furthermore, the cell detachment device 10 arranges the transducers 11 along an arbitrary array axis direction. Therefore, the cell detachment device 10 can reduce the cost since the ultrasonic probe 12 can be made smaller (the number of transducers 11 can be reduced) compared to a case where the transducers 11 are provided to cover the entire bottom surface of the culture vessel 2, for example.

[0030] (Culture vessel and cell adhesion surface) The culture vessel 2 has cells 1 therein. In this specification, the surface inside the culture vessel 2 to which the cells 1 are attached is referred to as the cell adhesion surface. The cell adhesion surface may be the inner wall of the culture vessel 2. Alternatively, another member may be placed inside the culture vessel 2, and the cells 1 may be cultured on the surface of that member. In this case, the surface of the member to which the cells 1 are attached becomes the cell adhesion surface.

[0031] The cell detachment device 10 is configured to irradiate an ultrasonic beam 13 onto cells 1 attached to a cell adhesion surface inside a culture vessel 2 and immersed in a culture solution 3 such as a medium, thereby detaching the cells 1 from the cell adhesion surface. The cells 1 are attached to the inner bottom surface of the culture vessel 2 in the form of, for example, cells, colonies, cell sheets, organoids, etc. That is, in this embodiment, the inner bottom surface of the culture vessel 2 is the cell adhesion surface.

[0032] 1A, ultrasonic beam 13 is indicated by two dotted lines. The entire area between the two dotted lines is ultrasonic beam 13. Ultrasonic beam 13 is emitted from transducer 11, passes through fluid 4 and the wall of culture vessel 2, and is irradiated to cells 1 in culture vessel 2. Since the present invention uses the acoustic force of ultrasound to detach cells 1, the energy of ultrasonic beam 13 can be transmitted between transducer 11 and culture vessel 2 even via fluid 4.

[0033] In FIG. 1A, the cells 1 are illustrated as a group of many cells 1. To give a specific example, the width of the transducer 11 in the long axis direction (y-axis in FIG. 1A) is typically 5 to 40 mm. Meanwhile, the size of each cell 1 is typically 10 to 50 micrometers. Therefore, many cells 1 are located within the range of the long axis width of the transducer 11. For convenience, in FIG. 1A, the many cells 1 are illustrated as a single oval. This is the same in other drawings in this specification.

[0034] The culture vessel 2 is partially immersed in a fluid 4, which is a contact medium in a tank 5 as shown in Fig. 1A and Fig. 1B. Specifically, as shown in Fig. 1A and Fig. 1B, a highly viscous fluid 4 that can efficiently transmit ultrasound can be supplied between the ultrasonic probe 12 and the culture vessel 2. In this embodiment, water can be used as such a fluid 4. Note that the configuration between the culture vessel 2 and the cell detachment device 10 is not limited to this.

[0035] 1A, an ultrasonic probe 12 is disposed outside the culture vessel 2 and is preferably immersed in the fluid 4. A transducer 11 in the ultrasonic probe 12 irradiates an ultrasonic beam 13 to a cell 1 in the culture vessel 2 through the fluid 4 and the cell adhesion surface.

[0036] (Ultrasound probe configuration) A configuration example of the transducer 11 (ultrasonic transducer) will be described with reference to Figs. 2A and 2B. Fig. 2A is a schematic cross-sectional view of the transducer 11. Fig. 2B is a schematic plan view of the transducer 11. As shown in Figs. 2A and 2B, the transducer 11 has, as an example, a flat-plate-shaped piezoelectric element 411. The transducer 11 has a ground electrode 412 on the upper surface side of the piezoelectric element 411, i.e., on the culture vessel 2 side, and has a protective film 415 bonded to the upper surface side of the ground electrode 412. The protective film 415 of the transducer 11 comes into contact with the fluid 4. The protective film 415 is a thin film made of metal or resin. Furthermore, the transducer 11 has a signal electrode 413 for applying a voltage to the lower surface of the piezoelectric element 411. The shape of the piezoelectric element 411 may be, but is not limited to, a circle, an ellipse, a rectangle (square in plan view in FIG. 2A), etc. The connection portion between the signal electrode 413 and the signal line 17 (FIG. 1A) is covered with resin and fixed.

[0037] The piezoelectric element 411 is an element that converts an electric signal into mechanical motion. In this embodiment, the piezoelectric element 411 emits an ultrasonic beam 13 into the fluid 4 by applying a high-frequency electric signal. The piezoelectric element 411 is made of, for example, ceramics such as lead zirconate titanate or lead titanate, materials such as zinc oxide or lithium niobate, or a composite (composite material). The ground electrode 412 and the signal electrode 413 are made of, for example, gold, silver, copper, platinum, titanium, aluminum, or other materials. The piezoelectric element 411 may be a composite piezoelectric material in which a number of piezoelectric materials are solidified with resin or the like.

[0038] 2B, the signal electrode 413 is shown by a dotted line because it is located on the back surface of the piezoelectric element 411. When a voltage is applied to the signal electrode 413, an electric field is applied to the piezoelectric element 411 in the region where the signal electrode 413 and the ground electrode 412 overlap, and an ultrasonic beam 13 is emitted from the overlapping region of both electrodes. In this way, by appropriately designing the overlapping shape of the signal electrode 413 and the ground electrode 412, a desired beam shape 310 (FIGS. 3A and 3B), such as a rectangle or an ellipse, can be obtained.

[0039] In this embodiment, the upper surface side is the ground electrode 412, but the upper surface side may be the signal electrode 413 and the lower surface side may be the ground electrode 412. Also, a voltage may be applied to both the ground electrode 412 and the signal electrode 413. In either case, it goes without saying that an appropriate electric field can be applied to the piezoelectric element 411, and therefore the ultrasonic beam 13 can be generated.

[0040] A plurality of transducers 11 may be configured for one piezoelectric element 411. As described above, the ultrasonic beam 13 is emitted from the overlapping region between the signal electrode 413 and the ground electrode 412. Therefore, by providing a plurality of signal electrodes 413, a plurality of overlapping regions between the signal electrode 413 and the ground electrode 412 can be created. Each overlapping region becomes a transducer 11. Even in this way, the transducer 11 that emits the ultrasonic beam 13 can be selected by selecting the signal electrode 413 to which a voltage is applied, so that the effect of this embodiment can be obtained.

[0041] (Cell surface and beam shape) Before describing the beam shape 310, the cell surface is defined. The cell surface is defined as the surface to which the cell 1 to be detached is attached in the culture vessel 2. Usually, the cell surface coincides with the cell adhesion surface. In this specification, the z-direction position of the cell surface is referred to as the cell surface height zc.

[0042] Next, the beam shape 310 of the ultrasonic beam 13 emitted from the ultrasonic probe 12 will be described with reference to Figures 3A and 3B. Figure 3A is a schematic diagram showing an example of the beam shape 310 of the ultrasonic beam 13. Figure 3B is a schematic diagram showing another example of the beam shape 310 of the ultrasonic beam 13. In this specification, the beam shape 310 is defined as the shape of the ultrasonic beam 13 on the cell surface.

[0043] As shown in Figures 3A and 3B, the beam shape 310 of the ultrasonic beam 13 emitted from the ultrasonic probe 12 has a flat shape. In this specification, a flat shape means a shape having different widths in directions perpendicular to each other, and includes, for example, a rectangle (specifically a rectangle in Figure 14A) and an ellipse (Figure 14B). In this specification, the longitudinal direction of the flat beam shape 310 is called the major axis 321, and the short side direction is called the minor axis 322. The minor axis 322 is set in a direction perpendicular to the major axis 321.

[0044] In this embodiment, the direction of the short axis 322 is usually set as the scanning axis direction (see FIG. 1B). The array axis along which the ultrasonic probes 12 are arranged is usually set as the direction of the long axis 321. When the ultrasonic probes 12 are installed in such a direction, the beam irradiation area becomes large when the ultrasonic beam 13 is scanned over a certain distance Δx, so that the cell 1 can be efficiently peeled off.

[0045] Generally, the long axis 321 is arranged perpendicular to the scanning axis, but it may be approximately perpendicular. In Fig. 3A, consider a case where the scanning axis of the ultrasonic beam 13 is the x-axis direction, and the long axis 321 of the beam shape 310 is shifted at an angle θ from the perpendicular direction (i.e., the y-axis). The width of the long axis 321 of the beam shape 310 is WL, and the sweep area ΔS when the beam shape 310 is scanned a distance Δx is given by Equation 1. ΔS=WL×Δx×cosθ (Formula 1)

[0046] ΔS given by formula 1 is maximum when θ=0, which is preferable since it allows efficient detachment of cell 1. However, even if θ deviates slightly, for example, when θ=40°, ΔS maintains 77% of the maximum value, and the effect of this embodiment can be obtained. In other words, even an angle deviated by ±40° from the y-axis can be regarded as "approximately right angles."

[0047] As a configuration for realizing the flat shape of the beam shape 310, for example, an overlapping region of electrode shapes sandwiching the piezoelectric element 411 may be flattened. Alternatively, the beam shape 310 may be flattened by selecting a specific transducer 11 from a plurality of array-shaped transducers 11. The configuration for making the beam shape 310 flat is not limited to this.

[0048] Alternatively, the transducer 11 may be curved so that its cross section is U-shaped, and the transducer 11 may emit a linearly converged ultrasonic beam 13. When such a linear convergence type ultrasonic probe is used, the beam emission area becomes wider, and therefore, an intense ultrasonic beam 13 can be obtained near the convergence position. Even when a linear convergence type ultrasonic probe is used, the beam shape 310 on the cell surface becomes flat. In this case, the long axis 321 direction and the short axis 322 direction are defined by the beam shape 310 on the cell surface.

[0049] (Relationship between the long axis of the flat shape and the scanning axis) In this embodiment, it is important that the beam shape 310 of the ultrasonic waves emitted by the transducer 11 is flattened, and that the direction substantially perpendicular to the major axis 321 of the beam is set as the scanning axis. The effect of this configuration will be described. Fig. 4A is an explanatory diagram showing the arrangement of the direction of long axis 321 of flat beam shape 310 and the scanning axis, and is a diagram showing a configuration in which the direction perpendicular to long axis 321 is the scanning axis. Fig. 4B is an explanatory diagram showing the arrangement of the direction of long axis 321 of flat beam shape 310 and the scanning axis, and is a diagram showing a configuration in which the direction of long axis 321 is the scanning axis. The width of long axis 321 is designated as WL, and the width of short axis 322 is designated as WS. Hereinafter, the configuration in Fig. 4A is referred to as configuration A, and the configuration in Fig. 4B is referred to as configuration B.

[0050] As an example, configuration A in FIG. 4A has WL=20 mm and WS=3 mm. Configuration B in FIG. 4B has WL=10 mm and WS=6 mm. The beam area of ​​beam shape 310 is WL×WS, so the beam area is equal in configurations A and B. Therefore, if the ease of detachment from the cell adhesion surface is constant regardless of location, the amount that can be detached by irradiating ultrasonic beam 13 once should be the same in both cases. However, when detaching cell 1, the following effects can be obtained by using configuration A.

[0051] The inventors have experimentally found that when cells 1 are detached by irradiation with an ultrasonic beam 13, the adhesiveness between the cells 1 and the cell adhesive surface varies depending on the location. For example, taking a cell sheet as an example of the cells 1, the cells 1 are easily detached by the ultrasonic beam 13 from the edge of the cell sheet, but are difficult to detach from the inner region of the cell sheet.

[0052] Furthermore, the inventors have found that the effect on the cells 1 is smaller when the cell sheet is peeled off from the edge. The reason for this will be described with reference to Figs. 5A and 5B. Fig. 5A is an explanatory diagram of the case where an ultrasonic beam 13 is irradiated to the inner region of a cell sheet adhered to the cell adhesion surface inside a culture vessel 2. Fig. 5A shows an enlarged view of the irradiation area of ​​the ultrasonic beam 13, and the culture solution 3 is omitted. As shown in Fig. 5A, when the ultrasonic beam 13 is irradiated, the cells 1-1 in the irradiated area are peeled off, but since the cells 1-1 in the irradiated area are in the inner region of the cell sheet, the cells 1 (cells 1-2 at the peeling boundary) around the peeled part remain attached to the cell adhesion surface. Therefore, the cells 1-2 at the peeling boundary are subjected to a strain stress that deforms the cells 1, which has a negative effect on the cells 1-2 at the peeling boundary. In other words, the cells 1-2 at the peeling boundary are subjected to mechanical stress, which has a negative effect on the cells 1-2 at the peeling boundary. In addition, the cell sheet may be damaged by the stress that mechanically stretches the cells 1-1 in the irradiated area.

[0053] 5B is an explanatory diagram of the case where the edge of the cell sheet is irradiated with an ultrasonic beam 13. In this case, the cell 1-1 in the irradiated part of the detached edge is not fixed to the cell adhesion surface, so the distortion stress on the cell 1-2 at the detachment boundary is significantly reduced. According to the inventors' experiments, what is considered to be the edge is within 5 mm from the edge of the cell sheet, and more preferably within 3 mm from the edge.

[0054] Returning to FIG. 4, the explanation continues. In the configuration A of FIG. 4A, since WS=3 mm, only the edge of the cell sheet is irradiated with one ultrasonic beam 13 irradiation. Therefore, the cell 1 is easily peeled off, and the strain stress on the cell 1 is at a negligible level. When the edge of the cell sheet is peeled off, the position of the peeling boundary becomes a new edge. Therefore, if the beam irradiation position is moved by the width WS=3 mm of the short axis 322 in the scanning axis direction and the next ultrasonic beam 13 is irradiated, the ultrasonic beam 13 is sequentially irradiated to a new edge region, so that the cell 1 can be easily peeled off. In this way, by using the flat beam shape 310 and scanning in a direction approximately perpendicular to the long axis 321 of the beam shape 310, the cell sheet can be peeled off sequentially from the edge, and the cell 1 can be easily peeled off.

[0055] On the other hand, in the configuration B of Fig. 4B, the ultrasonic beam 13 is irradiated from the edge of the cell sheet to a range of WL = 10 mm in one irradiation. Therefore, the ultrasonic beam 13 is also irradiated to the inner region of the cell sheet. Therefore, the cells 1 are difficult to peel off, and the strain stress on the cells 1 in the inner region increases. For the above reasons, in this embodiment, the width WS of the minor axis 322 of the beam shape 310 of the flat ultrasonic beam 13 is preferably 5 mm or less. In this embodiment, it is more preferable to set the width WS of the minor axis 322 to 3 mm or less. Furthermore, in this embodiment, it is even more preferable to set the width WS of the minor axis 322 to 2 mm.

[0056] In the above explanation, a cell sheet is used as an example, but the same applies to cells 1 that are not in a sheet form. Moreover, the configuration of this embodiment shown in FIG. 4A is effective in terms of high speed peeling and device costs. The drive current required for element driving section 14 that drives oscillator 11 is proportional to the area of ​​oscillator 11. As the drive current increases, the cost of the circuit that constitutes element driving section 14 increases, so the area of ​​oscillator 11 affects the cost. On the other hand, for high-speed detachment, it is effective to reduce the number of scans in the y-axis direction, so it is preferable to increase the width in the y-axis direction of transducer 11 (width WL of major axis 321). More preferably, the length in the y-axis direction of ultrasonic probe 12 is longer than the length in the y-axis direction of culture vessel 2. In this way, cells 1 in the entire culture vessel 2 can be detached by scanning only in the x-axis direction, so a one-axis scanning device is sufficient, which not only increases the speed but also reduces the cost of the device.

[0057] 4A (Configuration A) and 4B (Configuration B), when the area of ​​the transducer 11 is constant, the length of the ultrasonic probe 12 in the y-axis direction can be made longer in Configuration A. This provides the advantage of achieving both high speed peeling and low equipment costs. As described above, in the cell detachment device 10, the beam shape 310 of the ultrasonic beam 13 emitted by the transducer 11 is made flat, and the scanning axis is arranged roughly perpendicular to the long axis 321 of the beam shape 310, thereby achieving effects such as making it easier to detach the cells 1.

[0058] (Element drive unit and scanning mechanism) Let us return to Figure 1 to continue the explanation. The element driving unit 14 applies a voltage to the transducer 11 of the ultrasonic probe 12 based on a control command from the control unit 16, causing the transducer 11 to generate an ultrasonic beam 13 having a desired beam output. The element driving unit 14 is provided for each transducer 11, and applies a voltage to the transducer 11 independently. The voltage waveforms applied to the transducers 11 will be described later. The order in which the four transducers 11 shown in Figs. 1A and 1B are driven (the driving sequence) will also be described later.

[0059] The scanning mechanism 15 is a mechanism for relatively moving the ultrasonic probe 12 and the culture vessel 2. In this embodiment, the scanning mechanism 15 has a configuration for moving the ultrasonic probe 12 along the outer surface of the bottom of the culture vessel 2 and along a trajectory calculated by the control unit 16. A well-known configuration such as an XY-axis moving table can be used as the configuration of the scanning mechanism 15. Note that the scanning mechanism 15 may be attached to the culture vessel 2 and move the positions of the ultrasonic probe 12 and the culture vessel 2 relatively by scanning the culture vessel 2.

[0060] (Control unit and scanning path calculation unit) The configuration of the control unit 16 will be described with reference to Fig. 6. Fig. 6 is a schematic diagram of the cell detachment device 10, in which the control unit 16 is explained in detail. As shown in Fig. 6, the control unit 16 has a beam control unit 53, a beam condition determination unit 54, a scan path calculation unit 55, and a planning unit 59 as internal functions embodied by executing software on hardware. In the example shown in Fig. 6, the beam control unit 53 is connected to each element driving unit 14 and controls the driving of each element driving unit 14. The scan path calculation unit 55 is connected to the scanning mechanism 15 and controls the scanning by the scanning mechanism 15.

[0061] In other words, the beam control unit 53 and the scanning path calculation unit 55 calculate the path for moving the ultrasonic probe 12 to peel off the desired area of ​​the cell 1 and the output of the ultrasonic beam 13 at a position on the path, and control the scanning mechanism 15 and the ultrasonic probe 12.

[0062] For example, the planning unit 59 refers to the irradiation conditions of the ultrasonic beam 13 inputted by the input unit 56 and the coordinates of the desired ablation region 511, and plans the intensity of the ultrasonic beam 13 and the coordinates to be scanned. Based on this, the scanning path calculation unit 55 issues a scanning instruction to the scanning mechanism 15.

[0063] The beam condition determination unit 54 determines the voltage and waveform to be applied to the transducer 11 by the element driving unit 14 based on the irradiation conditions of the ultrasonic beam 13 planned by the planning unit 59. The transducer 11 that irradiates ultrasonic waves may be switched on software using a circuit based on an electric signal. Also, the transducer 11 that irradiates ultrasonic waves may be switched by a physical button or the like.

[0064] The beam control unit 53 controls the conditions of the voltage to be applied to the element driving unit 14 based on the applied voltage and waveform determined by the beam condition determination unit 54. The beam control unit 53 calculates the signal and intensity of the voltage to be applied to the ultrasonic probe 12.

[0065] The scanning path calculation unit 55 calculates the path of the ultrasonic probe 12 moved by the scanning mechanism 15 based on the irradiation position planned by the planning unit 59. The trajectory of the ultrasonic beam 13 by the scanning path calculation unit 55 can be arbitrarily set by the input unit 56. The scanning path calculation unit 55 calculates the coordinates of the ultrasonic probe 12 based on the input shape so that the ultrasonic beam 13 is irradiated along the input shape, starting from an arbitrary coordinate point of an arbitrary ultrasonic probe 12. After irradiating the planned irradiation point with the ultrasonic beam 13, the scanning path calculation unit 55 calculates the movement amount of the ultrasonic probe 12 to the next specified position.

[0066] The input unit 56 inputs information specifying a region to be detached from the cells 1 on the cell adhesion surface of the culture vessel 2. For example, the input unit 56 can input information for cutting out a detached portion having a desired shape from the cells 1 being cultured displayed on the display unit 57. The input unit 56 has a keyboard and a mouse, and an operator or the like inputs information, and the input information such as the shape of the detached portion is provided to the database 52 and the control unit 16.

[0067] The display unit 57 is provided to assist the input unit 56, and allows input while referring to input information. The display unit 57 may be provided as necessary. The database 52 stores the irradiation conditions of the ultrasonic beam 13, i.e., the control data of the ultrasound required for detaching the cells 1, such as the size, intensity, and transmission time of the ultrasonic beam 13, depending on the type of ultrasonic probe 12 (or transducer 11) used, the type of cultured cells, the type of culture vessel 2, the number of days of culture, the state of culture, etc. Although the database 52 is not required, providing the database 52 enables the control unit 16 (e.g., the beam condition determination unit 54) to appropriately set the irradiation conditions of the ultrasonic beam 13 according to the type and state of the cell 1, the type of culture vessel 2, etc., thereby enabling the cell 1 to be detached more efficiently.

[0068] (Vibrator drive waveform) FIG. 7 is an explanatory diagram illustrating an example of a voltage waveform output from the element driving unit 14 and applied to each transducer 11 constituting the ultrasonic probe 12. As shown in FIG. The voltage waveform applied by the element driving section 14 is a burst waveform, and is composed of a wave packet made up of N fundamental waves. One wave packet is a waveform in which a waveform of a fundamental frequency f0 (fundamental wave) is repeated N cycles. In this specification, the period between a wave packet and the next wave packet is called a repetition period Tr. In this embodiment, the repetition period Tr is typically 0.2 seconds to 2 seconds. In this embodiment, it is preferable to set the repetition period Tr to 1 second. The repetition period Tr of the wave packet is not limited to these values.

[0069] In this embodiment, the fundamental wave can be a sine wave, but a square wave or a triangular wave may also be used, as shown in Fig. 7. Using a square wave has the advantage that the circuit configuration of the element driving unit 14 is simplified. In this embodiment, the fundamental frequency f0 can be set to 1 MHz, and the number of cycles N of the fundamental wave can be set to 3000. In other words, N cycles of the fundamental wave constitute one wave packet. It goes without saying that the setting ranges of the fundamental frequency f0 and the number of cycles N are not limited to these.

[0070] The fundamental frequency f0 can be set to 150 kHz or more and 50 MHz or less. It is known that if the fundamental frequency f0 is set to less than 150 kHz, cavitation is more likely to occur in the culture fluid 3, although this depends on the type of culture fluid 3, etc. Shock waves emitted by cavitation may affect the cells 1, so it is desirable to set the fundamental frequency f0 to 150 kHz or more, and more desirable to set the fundamental frequency f0 to 200 kHz or more so as not to affect the cells 1. Furthermore, if the frequency exceeds 50 MHz, it becomes difficult to design the element driving unit 14. It is more preferable that the range of the fundamental frequency f0 is 400 kHz or more and 5 MHz or less. It should be noted that one period T 0 is the fundamental frequency f0 divided by 1 (i.e., T 0 =1 / f0).

[0071] The wavelength of ultrasound is expressed by Equation 2, where c is the speed of sound (m / s) and f is the frequency (Hz). λ=c / f (Equation 2) Since the speed of sound in the culture solution 3 is approximately 1500 (m / s), when f0=1 MHz, the wavelength λ is 1.5 mm. In this embodiment, as described above, the acoustic force of the ultrasonic beam 13 is used, so if the ultrasonic beam 13 spreads, the acoustic force per unit area at the cell adhesion surface decreases. In order to keep the spread of the ultrasonic beam 13 to about a few mm, it is preferable to set the ultrasonic wavelength to 4 mm or less. To realize this, it is preferable that the fundamental frequency f0 is 400 kHz or more.

[0072] (Vibrator drive sequence) Next, the driving sequence of the multiple transducers 11 in the cell detachment device 10 shown in FIG. 1 will be described. FIG. 8 is a schematic diagram showing the cells 1 on the cell culture surface in the culture vessel 2 in a detachment region 511 and a non-detachment region 512. In the figure, the region shown with diagonal lines is the non-detachment region 512, and the other region is the detachment region 511. The numbers 1 to 4 arranged vertically correspond to the numbers of the four transducers 11. The characters t1, t2,... arranged horizontally indicate the x-direction positions at which the cells 1 are detached at times t1 and t2, respectively. As shown in FIG. 8, in this embodiment, the element driving unit 14 is turned on / off and the output is controlled for each of the four transducers 11 from the left end of the culture vessel 2 in response to times t1, t2,... This allows control to be performed such that the cells 1 are detached in the detachment region 511 and the cells 1 are not detached in the non-detachment region 512.

[0073] FIG. 9 is a schematic diagram showing an example of time-series control of the transducer 11 in the first embodiment. Fig. 9 shows a drive sequence of the transducer 11 for detaching the cell 1 in the detachment region 511 in Fig. 8. The numbers 1 to 4 in the vertical direction in Fig. 9 correspond to the numbers of the four transducers 11. In the sequence in Fig. 9, the horizontal axis represents elapsed time, and shows the sequence up to time t2 shown in Fig. 8.

[0074] In each waveform, a low level represents a state in which no voltage is applied to the ultrasonic probe 12, and a high level represents a wave packet to be applied. Here, the wave packet is as described above with reference to FIG. As shown in FIG. 8, at time t1, cells 1 in all regions corresponding to transducers 11 numbers 1 to 4 are to be detached, and therefore, as shown in FIG. 9, wave packets are applied to four transducers 11 at time t1.

[0075] Next, the transducer 11 is scanned in the x-axis direction by the scanning mechanism 15. After that, at time t2, the cells 1 in all areas corresponding to the transducers 11 numbered 1 and 4 are peeled off, so as shown in Fig. 9, at time t2, the wave packet is applied only to the two transducers 11 numbered 1 and 4. Although the sequence after time t3 is omitted, in this manner, the cells 1 can be selectively detached from a desired detachment region 511 as shown in Fig. 8. In this manner, according to this embodiment, by using a plurality of transducers 11, the cells 1 can be selectively detached from a desired region of the cell adhesion surface in the culture vessel 2.

[0076] In this way, the cell detachment device 10 according to this embodiment can irradiate the cells 1 being cultured with the ultrasonic beam 13, thereby efficiently detaching the cells 1 in any shape and in a non-contact manner from the culture vessel 2. In particular, the cell detachment device 10 according to this embodiment independently controls and scans the multiple transducers 11 aligned in the array axis direction with respect to the desired detachment region 511, thereby expanding the region irradiated with the ultrasonic beam 13 and enabling faster detachment.

[0077] (Transducer placement) Here, the arrangement of the multiple transducers 11 will be described. Figures 10A to 10D are schematic diagrams for explaining an example of the arrangement axis and scanning axis of the transducers 11. All of Figures 10A to 10D show the arrangement of the multiple transducers 11. For the sake of simplicity, Figures 10A to 10D show a configuration using three transducers 11.

[0078] 10A shows a configuration in which the transducer 11 is arranged so that the long axis direction of the beam shape 310 of the ultrasonic beam 13 emitted from the transducer 11 on the cell adhesion surface is aligned with the array axis. The scanning axis along which the transducer 11 scans is perpendicular to the array axis. In other words, the beam shape 310 of the ultrasonic beam 13 emitted from the transducer 11 is flat, and the transducer 11 is arranged so that the long axis of the flat shape is perpendicular to the scanning axis.

[0079] 10B shows a configuration in which the scanning axis is approximately perpendicular to the array axis. In other words, the beam shape 310 of the ultrasonic beam 13 emitted by the transducer 11 is flat, and the transducer is arranged so that the long axis of the flat shape is approximately perpendicular to the scanning axis. When the array axis is tilted by θ from the direction perpendicular to the scanning axis (y axis), as described above, if the absolute value of θ is 40° or less, the rate of decrease in the cell detachment sweep area ΔS expressed by Equation 1 is small, and therefore the array axis can be considered to be "approximately perpendicular."

[0080] 10C shows a configuration in which adjacent transducers 11 are arranged to be shifted from each other in the direction of minor axis 322 of flat beam shape 310, and arranged to overlap in the direction of major axis 321. That is, in FIG. 10C, adjacent transducers 11 are arranged so that the overlap amount Lov of the positions in the direction of major axis 321 is positive (i.e., so as to overlap). Arranging in this manner is more preferable, as it eliminates peeling residue in adjacent transducers 11.

[0081] Fig. 10D shows a modified example of the configuration shown in Fig. 10C. Fig. 10D shows a configuration in which adjacent transducers 11 are arranged to be offset in the direction of minor axis 322 of flat beam shape 310, and arranged to overlap in the direction of major axis 321. That is, also in Fig. 10D, adjacent transducers 11 are arranged so that the overlap amount Lov of the positions in the direction of major axis 321 is positive. With such an arrangement, it is possible to obtain the same effect as the configuration shown in Fig. 10C. The arrangement of transducers 11 is not limited to the configurations shown in FIGS. 10A to 10D.

[0082] (fluid) 1, in this embodiment, a fluid 4 is placed inside a tank 5, and a culture vessel 2 is placed in the fluid 4. In this manner, an ultrasonic beam 13 emitted from a transducer 11 constituting an ultrasonic probe 12 is irradiated to the culture vessel 2 through the fluid 4. By adopting a configuration in which the ultrasonic beam 13 is irradiated to the culture vessel 2 through the fluid 4 in this manner, an effect is obtained in that scanning of the ultrasonic beam 13 is facilitated.

[0083] It is more preferable that the fluid 4 has a viscosity of 20 mPa·s or less, since this reduces the contact resistance during scanning. In this embodiment, water can be used as the fluid 4. The viscosity of water varies depending on the temperature, but is in the range of 0.7 to 1.3 mPa·s.

[0084] In this specification, the distance between the ultrasound emission surface of the transducer 11 and the culture vessel 2 is defined as the gap length Lg. The gap between the ultrasound emission surface and the culture vessel 2 is filled with the fluid 4. In order for the ultrasound beam 13 to be efficiently irradiated to the culture vessel 2, it is preferable that the gap length Lg is smaller than twice the wavelength λw of the ultrasound beam 13 in the fluid 4. That is, Lg≦2×λw (Formula 3) It is preferable to set the above. The wavelength λw of the ultrasonic beam 13 is determined by Equation 2. For example, when water is used as the fluid 4 and an ultrasonic beam 13 with a fundamental frequency of 1 MHz is used, the wavelength λw in the fluid 4 is 1.5 mm, so that the gap length Lg is preferably 3 mm or less. In this embodiment, Lg can be set to, for example, 1 mm.

[0085] (First Modification of the First Embodiment) Next, a cell detachment device 10 according to a first modified example of the first embodiment will be described. Fig. 11 is a schematic diagram of the cell detachment device 10 according to the first modified example of the first embodiment. Fig. 12 is a schematic diagram showing an example of the configuration of the cell detachment device 10 according to the first modified example of the first embodiment, and particularly explaining in detail an example of the configuration of the control unit 16. This modified example will be described mainly with respect to the differences from the first embodiment.

[0086] As shown in FIG. 11, in this modified example, there is one element driving unit 14 that applies voltage to the transducer 11, and the transducer 11 to be driven is selectively switched by a switching unit 31 to irradiate an ultrasonic beam 13, which is different from the first embodiment described above. 12, the present modification is different from the first embodiment in that the beam control unit 53 is connected to one element driving unit 14. The beam control unit 53 controls the condition of the voltage to be applied to this element driving unit 14 based on the applied voltage and waveform determined by the beam condition determination unit 54. The beam control unit 53 calculates the signal and intensity of the voltage to be applied to the ultrasonic probe 12. The switching unit 31 includes, for example, a relay circuit, and switches the transducers 11 that transmit the ultrasonic beams 13 determined by the control unit 16 in an arbitrary order, and applies a voltage to the transducers 11 one by one. In the cell dissection device 10 according to this modification, similarly to FIG. 1B, the ultrasonic probe 12 performs scanning in the direction of the minor axis 322 of the beam shape 310, that is, in the x-axis direction. According to this modification, the number of element driving sections 14 can be reduced, leading to lower costs.

[0087] Next, a description will be given of a drive sequence of the four transducers 11 in the cell detachment device 10 shown in Fig. 11 and Fig. 12. Fig. 13 is a schematic diagram showing an example of time-series control of the transducers 11 in the first modified example of the first embodiment. The drive sequence described here is a sequence for selectively detaching the cell 1 from the detachment region 511 shown in FIG.

[0088] As shown in FIG. 8, at time t1, cells 1 in all regions corresponding to transducers 11 numbered 1 to 4 are peeled off, so as shown in FIG. 13, wave packets are sequentially applied to transducers 11 numbered 1 to 4 from time t1.

[0089] Next, the transducer 11 is scanned in the x-axis direction by the scanning mechanism 15. After that, at time t2, the cells 1 in all regions corresponding to the transducers 11 numbered 1 and 4 are peeled off, so as shown in Fig. 13, from time t2, wave packets are applied sequentially to the transducers 11 numbered 1 and 4.

[0090] In this modified example, as an example, one wave packet is formed by a fundamental wave with a fundamental frequency of 1 MHz and a cycle number N=3000 times. Therefore, the output time of one wave packet is 3 ms. Including the switching time in the relay circuit, the delay times of the wave packets of adjacent ultrasonic probes 12 are shifted by 10 ms before output. Therefore, the time required to drive four transducers 11 is 33 ms. On the other hand, the time between time t1 and time t2 (repetition period Tr of the wave packet) is preferably set to 1 second. Thus, in a typical example, the output time of the wave packet is sufficiently short compared to the repetition period Tr of the wave packet, so that the required time is short even if the wave packets are output sequentially while switching the voltage, as shown in FIG. 13. Although the sequence after time t3 is omitted, in this manner, the cells 1 can be selectively detached from the desired detachment region 511 as shown in Fig. 8. In this manner, according to this modification, by using a plurality of transducers 11, the cells 1 can be selectively detached from the desired region of the cell adhesion surface in the culture vessel 2. In this way, in the cell peeling device 10 according to this modified example, the number of element driving sections 14 can be reduced, leading to lower costs.

[0091] (Second Modification of the First Embodiment) Next, a cell detachment device 10 according to a second modified example of the first embodiment will be described. Fig. 14 is a schematic configuration diagram of the cell detachment device 10 according to the second modified example of the first embodiment. Note that this modified example will be described mainly with respect to the differences from the first embodiment.

[0092] 14, this modification is different from the first embodiment in that a beam variable mechanism 41 is provided on the ultrasonic probe 12 and the beam shape 310 of the ultrasonic beam 13 can be varied. The cell detachment device 10 according to this modification can adjust the intensity of the irradiated ultrasonic beam 13 by the beam variable mechanism 41. That is, the cell detachment device 10 according to this modification can irradiate the ultrasonic beam 13 with localized intensity changes by the beam variable mechanism 41 in response to adhesion states that differ depending on the adhesion degree of the cells 1. Therefore, the cell detachment device 10 according to this modification can efficiently detach the cells 1.

[0093] The beam varying mechanism 41 may be provided corresponding to a plurality of transducers 11 constituting the ultrasonic probe 12. In this way, each transducer 11 can vary the beam shape 310, so that the cells 1 can be detached more finely. Furthermore, when the beam variable mechanism 41 is shared by a plurality of transducers 11, one beam variable mechanism 41 may be provided in the ultrasonic probe 12.

[0094] The beam variable mechanism 41 can be composed of, for example, one or more acoustic lenses. The acoustic lens is controlled by the control unit 16 to change the focal position in the transmission axis direction of the ultrasonic beam 13. The beam shape 310 of the ultrasonic beam 13 on the cell adhesion surface is flat with the major axis in the y-axis direction. In this modification, the beam shape 310 of the ultrasonic beam 13 on the cell adhesion surface is set to a rectangle. In the cell detachment device 10 according to this modification, similar to FIG. 1B, the ultrasonic probe 12 also scans in the direction of the minor axis 322 of the beam shape 310, that is, in the x-axis direction. With such a configuration, the cell detachment device 10 according to this modification can efficiently detach the cell 1.

[0095] The output conditions of the ultrasonic beam 13, including the focusing conditions of the ultrasonic waves by the acoustic lens, are determined by the control unit 16. In this modification, a Fresnel lens for changing the focus of the ultrasonic beam 13 may be provided, and a voltage may be applied to the piezoelectric element 411 (see FIG. 2A, etc.) of the transducer 11 by modulating the frequency. In this case, the control unit 16 may determine the output conditions of the ultrasonic beam 13, including the Fresnel lens and the modulated frequency. In this manner, the cell detachment device 10 according to this modified example can vary the intensity of the ultrasonic beam 13 locally, thereby enabling the cells 1 to be detached efficiently.

[0096] Second embodiment Next, a cell detachment device 10 and a cell detachment method according to a second embodiment will be described. Fig. 15 is a schematic diagram of a cell detachment device 10 according to the second embodiment. Fig. 16 is a schematic diagram showing an example of a database 52 having ultrasonic irradiation conditions. The present embodiment will be described mainly with respect to the differences from the first embodiment.

[0097] 15, the cell detachment device 10 according to the second embodiment differs from the configuration of the first embodiment in that it additionally includes a detection unit 51 that detects the detached state (adhesion state) of the cell 1. The cell detachment device 10 according to the second embodiment also differs from the configuration of the first embodiment in that it includes a recognition unit 58 instead of the planning unit 59 in the first embodiment. This recognition unit 58 recognizes whether or not the cell 1 has been detached based on the information detected by the detection unit 51. In addition, the cell detachment device 10 of the second embodiment, like the first embodiment (see Figure 6), is equipped with a database 52, a beam control unit 53, a beam condition determination unit 54, a scanning path calculation unit 55, an input unit 56, and a display unit 57.

[0098] Through various investigations, the present inventors have found that there is variation (unevenness) in the adhesion state of the cells 1. That is, the present inventors have found that there is a difference in adhesive force between the cells 1 and the cell adhesion surface depending on the location of the cell adhesion surface in the culture vessel 2. Therefore, depending on the intensity of the ultrasonic beam 13, after irradiation with one wave packet of the ultrasonic beam 13, the cells 1 may be divided into those that are detached and those that remain attached without being detached.

[0099] In the second embodiment, based on the fact experimentally confirmed that there is variation (unevenness) in the adhesion state of the cells 1, the presence or absence of detachment of the cells 1 is checked for each irradiation of the ultrasonic beam 13, and if the cells 1 are not detached, the ultrasonic beam 13 is irradiated again. In this way, in the second embodiment, it is possible to realize detachment of the cells 1 without leaving any detached cells 1 and with the irradiated ultrasonic energy reduced as much as possible. In the second embodiment, since the intensity of the ultrasonic beam 13 can be reduced, the cells 1 can be detached without affecting them.

[0100] Note that each irradiation of the ultrasonic beam 13 means each irradiation of a wave packet of a burst wave. That is, in the second embodiment, after one wave packet is irradiated, the state of the cell 1 is detected by a detection unit 51 and a recognition unit 58 described later, and the next ultrasonic irradiation position is determined.

[0101] The detection unit 51 detects the detached state of the cells 1. The detached state refers to the state of how many cells 1 have detached within a given area, and means, for example, the proportion of detached cells 1 within a given area. The proportion of cells 1 in an adherent state is also an index of the detached state. Since the proportion of adhered cells 1 (cells 1 in an adherent state) can be detected and converted into a detached state, the detached state may be detected by detecting the adherent state.

[0102] For example, an optical camera or an optical microscope can be used as the detection unit 51. The detection unit 51 may measure data such as the shape of the cell 1 and the coordinates of the end portion, and store the data in the database 52. The recognition unit 58 recognizes the shape and detached state of the cell 1. For recognition, image processing using an image of the observed cell 1 may be used. Here, for example, the detached portion of the cell 1 is disturbed by the acoustic force of the ultrasonic beam 13, but the portion of the cell 1 adhering to the cell adhesion surface is not disturbed even when it is subjected to the acoustic force of the ultrasonic beam 13. Therefore, the detached state of the cell 1 can be recognized by observing these conditions.

[0103] In the study, the inventors used an optical camera as the detection unit 51. The optical camera was installed on the upper side of the culture vessel 2. That is, the inventors installed the optical camera, which is the detection unit 51, on the opposite side to the side where the ultrasonic beam 13 is irradiated. By installing it on the opposite side, there is an effect that there is no physical interference between the ultrasonic probe 12 and the optical camera, which is the detection unit 51. In the second embodiment, images before and after irradiation of the wave packet of the ultrasonic beam 13 are sent to the recognition unit 58. The recognition unit 58 detects the detachment state of the cells 1 by comparing the images before and after irradiation. That is, the recognition unit 58 calculates the proportion of the cells 1 that remain attached without being detached (cells 1 that remain undetached).

[0104] In the second embodiment, as described above, an optical camera is used as the detection unit 51 as an example, but the remaining cells 1 may be detected by using the reflected signal of the ultrasonic beam 13. In this manner, the detection unit 51 may be any unit that detects the detached state of the cells 1. Note that in FIG. 15, the detection unit 51 is shown inside the control unit 16, but the detection unit 51 may be provided outside the control unit 16. That is, in the second embodiment, it is sufficient that the cell detachment device 10 is provided with the detection unit 51.

[0105] In the second embodiment, the steps (operations) to be performed as follows are appropriately set based on the proportion of the remaining cells 1 that have not been detached. First, the database 52 saves (stores) data on the shape (coordinate data) and adhesion state of the cell 1 recognized by the detection unit 51 and the recognition unit 58. The database 52 also stores irradiation conditions of the ultrasonic beam 13, that is, control data of the ultrasonic waves required for detaching the cell 1, such as the size, intensity, and transmission time of the ultrasonic beam 13, depending on the type of ultrasonic probe 12 (or transducer 11) used, the type of cultured cell, the number of days of culture, and the state of culture. For example, as shown in FIG. 16, the database 52 stores information such as the type AA of the ultrasonic probe 12, the type BB of the cultured cells, the number of days of culture CC, the applied voltage DD~, the wave number (number of sine waves) HH~ of the ultrasonic beam 13, the frequency LL~, and the number of irradiations per location PP~.

[0106] Next, based on the information in the database 52 , the beam condition determination unit 54 sets the basic conditions for the irradiation conditions of the ultrasonic beam 13 . The beam condition determination unit 54 determines the output of the ultrasonic beam 13 based on the irradiation conditions of the ultrasonic beam 13 stored in the database 52. Specifically, the beam condition determination unit 54 determines the voltage and waveform that the element driving unit 14 applies to the transducer 11. Here, the "output of the ultrasonic beam 13" includes the irradiation intensity of the ultrasonic beam 13, and a determination of whether or not to output the ultrasonic beam 13. Furthermore, the beam condition determination unit 54 may determine the conditions by referring to information on the detached state of the cells 1 obtained by the detection unit 51 and the recognition unit 58, as necessary. For example, if the first irradiation of the ultrasonic wave packet results in some cells 1 remaining after detachment, the irradiation intensity of the second wave packet may be increased. Alternatively, if the proportion of cells 1 remaining after detachment is small, the irradiation intensity of the second wave packet may be decreased compared to the first. Here, the irradiation intensity of the ultrasonic beam 13 is an amount determined by the voltage applied to the transducer 11, the number of cycles N that constitutes the wave packet, and the like.

[0107] The scanning path calculation unit 55 calculates a path for peeling off a desired peeling region 511 of the cell 1. The scanning path starts from any coordinate of any element number, and scans the entire surface of the peeling region 511 so as to fill it. In addition, the scanning path calculation unit 55 may scan the edge of the group of cells 1 detected by the detection unit 51 by irradiating it with an ultrasonic beam 13 to peel it off, and then irradiate the ultrasonic beam 13 to the boundary between the detached area 511 where the cells 1 have been detached and the adhesion area (area 512 that is not detached). The scanning path calculation unit 55 can refer to information on the detachment state of the cell 1 obtained by the detection unit 51. Specifically, if the cell 1 has not been sufficiently detached in the current beam irradiation area, the scanning path calculation unit 55 (i.e., the control unit 16) determines a scanning path to irradiate the same area with the ultrasonic beam 13 again. If the cell 1 has been sufficiently detached, the scanning path calculation unit 55 (i.e., the control unit 16) determines a scanning path to move to the next irradiation area.

[0108] If there is a cell 1 that has not been peeled off, the beam condition determination unit 54 and the scanning path calculation unit 55 may be configured so that only the corresponding transducer 11 among the plurality of transducers 11 irradiates the ultrasonic beam 13. For example, as shown in Fig. 9 and Fig. 13, after the first wave packet is irradiated at time t1, if the proportion of the cell adhesion surface that has not been peeled off is high on the cell adhesion surface corresponding to the transducers 11 numbered 1 and 4, the second wave packet at time t2 may be irradiated only by the transducers 11 numbered 1 and 4. In this embodiment, since a configuration using a plurality of transducers 11 is used, the amount of irradiation energy of the ultrasonic beam 13 can be reduced.

[0109] The input unit 56 inputs data (information) specifying a region to be detached from the cells 1 on the cell adhesion surface of the culture vessel 2. For example, the input unit 56 can input information for cutting out a detached portion having a desired shape from the cells 1 being cultured displayed on the display unit 57. The input unit 56 has a keyboard and a mouse, and is operated by an operator or the like to input information. The information such as the shape of the detached portion input from the input unit 56 is provided to the database 52 and the control unit 16.

[0110] The display unit 57 has a monitor screen and displays at least one of the actual image acquired by the detection unit 51, the detached area 511, the shape of the detached part of the cell 1 (the shape of the cell 1) inputted by the input unit 56, the mode of the ultrasonic waves being irradiated, the scanning path, and the irradiation position. In this way, the operator or the like can obtain appropriate information.

[0111] (Modification of the embodiment: Updating the database) In yet another embodiment, the control unit 16 includes an external connection unit (not shown) that enables the information in the database 52 to be updated from the outside. Here, the "external connection unit" may be, for example, a unit that directly connects an external storage device such as a hard disk drive, or may be a unit that connects to a network and performs updates from an external update database. This configuration is more preferable in that the information in the database 52 can be updated to the latest content.

[0112] (Cell detachment method) Next, one aspect of the cell detachment method according to the present embodiment will be described with reference to Fig. 17. Fig. 17 is a flow chart illustrating one aspect of the cell detachment method according to the present embodiment. As shown in FIG. 17, one aspect of the cell detachment method according to this embodiment includes a selection step S171, a scanning path calculation step S172, a movement step S173, and a detachment step S174.

[0113] In the selection step S171, a shape of the detached region 511 to be detached from the cell 1 is selected. The selection step S171 can be performed by the input unit 56. In the scanning path calculation step S172, a scanning path of the transducer 11 corresponding to the shape selected in the selection step S171 is calculated. The scanning path calculation step S172 can be performed by the scanning path calculation unit 55. In the moving step S173, the transducer 11 is moved to the designated position calculated and determined in the scanning path calculation step S172. The moving step S173 can be performed by the scanning mechanism 15. In the detachment step S174, a voltage is applied to the transducer 11, ultrasonic waves are generated from the transducer 11, and the ultrasonic beam 13 is irradiated to detach the cell 1 from the cell adhesion surface. The detachment step S174 can be performed by the beam condition determination unit 54, the beam control unit 53, the element driving unit 14, the ultrasonic probe 12, etc.

[0114] In one aspect of the cell detachment method according to the present embodiment, the transducers 11 are arranged along an arbitrary array axis direction (see, for example, FIGS. 10A to 10D). The beam shape 310 of the ultrasonic beam 13 emitted by the transducer 11 is flat, and the long axis 321 is arranged (approximately) perpendicular to the scanning axis. In addition, in the movement step S173, the scanning mechanism 15 relatively moves the position of the transducer 11 with respect to the culture vessel 2, causing the transducer 11 to scan along a scanning axis direction different from the array axis direction. As a result, in one aspect of the cell detachment method according to the present embodiment, as described in the cell detachment device 10, the beam irradiation area becomes large when the ultrasonic beam 13 is scanned over a certain distance, so that the cell 1 can be efficiently detached from the culture vessel 2 in a non-contact manner.

[0115] Next, another aspect of the cell detachment method according to the present embodiment will be described with reference to Fig. 18. Fig. 18 is a flow chart illustrating another aspect of the cell detachment method according to the present embodiment. As shown in FIG. 18, another aspect of the cell detachment method according to this embodiment includes a selection step S181, a scanning path calculation step S182, a movement step S183, a detachment step S184, a detection step S185, and an adjustment step S186.

[0116] Here, the selection step S181, the scanning path calculation step S182, the movement step S183, and the peeling step S184 in the other aspect are similar to the selection step S171, the scanning path calculation step S172, the movement step S173, and the peeling step S174 in the above-mentioned aspect, so the description thereof will be omitted and only the detection step S185 and the adjustment step S186 will be described.

[0117] In the detection step S185, the detached state of the cell 1 at the location irradiated in the detachment step S184 is detected. The detection step S185 can be performed by the detection unit 51 and the recognition unit 58. In the adjustment step S186, the output condition and irradiation position of the ultrasonic beam 13 for detaching the undetached cell 1 detected in the detection step S185 are adjusted. The adjustment step S186 can be performed by the beam condition determination unit 54 after detection by the detection unit 51 and the recognition unit 58, as described above.

[0118] In another aspect of the cell detachment method according to the present embodiment, the transducers 11 are arranged along an arbitrary array axis direction, as in the above-mentioned aspect (see, for example, FIGS. 10A to 10D). The ultrasonic beam 13 emitted by the transducer 11 has a flat beam shape 310, and its long axis 321 is arranged (almost) perpendicular to the scanning axis. In addition, in the moving step S183, the transducer 11 is caused to scan along a scanning axis direction different from the array axis direction by the scanning mechanism 15 that moves the position of the transducer 11 relative to the culture vessel 2. As a result, in the other aspect of the cell detachment method according to the present embodiment, as described in the cell detachment device 10, the beam irradiation area when the ultrasonic beam 13 is scanned over a certain distance is increased, so that the cell 1 can be efficiently detached from the culture vessel 2 without contact.

[0119] Next, a specific example of the cell detachment method according to this embodiment will be described with reference to Fig. 19. Fig. 19 is a flowchart for explaining the contents of a specific example of the cell detachment method according to this embodiment. Note that in the explanation of this specific example, multiple transducers 11 are scanned independently, but the scanning steps (movement steps S173, S183) are shown as a flowchart of the detachment method for any one ultrasonic probe 12. Note that the cell detachment method according to this embodiment is not limited to these, and is not limited to the following.

[0120] 19, first, the detachment conditions for the cell 1 are input to the input unit 56. Examples of the detachment conditions to be input include the selection of the area to be detached, the intensity of the ultrasonic beam 13 used for detachment, and the number of the transducer 11 to be used (step S191) (selection steps S171 and S181).

[0121] The scanning path calculation unit 55 reads out the coordinates of the arbitrary shape input in step S191 relative to the origin of the peeling portion (the origin is, for example, the center of the culture vessel 2) stored in the database 52. Next, the scanning path calculation unit 55 calculates the transducer 11 (ultrasonic probe 12) corresponding to the shape selected in step S191 and its scanning path (step S192) (scanning path calculation steps S172, S182).

[0122] Next, the scanning mechanism 15 changes the relative positions of the transducer 11 (ultrasonic probe 12) and the incubation container 2. Specifically, the scanning mechanism 15 moves the transducer 11 (ultrasonic probe 12) to a specified position so that the center of the ultrasonic beam 13 is positioned at the initial ultrasonic transmission point (origin) (step S193) (moving steps S173, S183).

[0123] Next, the beam condition determination unit 54 adjusts and determines the intensity and number of times of the ultrasonic beam 13 based on the ultrasonic beam 13 stored in the database 52 and the irradiation conditions inputted at the input unit 56 (step S194).

[0124] Next, the element driving unit 14 applies a voltage to the transducer 11 of the ultrasonic probe 12 to generate ultrasonic waves and irradiate the ultrasonic beam 13, thereby detaching the cell 1 (step S195) (detachment steps S174, S184).

[0125] Next, the detection unit 51 measures the cell 1 at the location irradiated with the ultrasonic beam 13. Based on the measured data, the recognition unit 58 recognizes the presence or absence of detachment (step S196) (detection step S185).

[0126] If no peeling has occurred (No in step S196), the process returns to step S194 where the intensity and number of times of the ultrasonic beam 13 are determined again by the beam condition determination unit 54 at the same coordinates (adjustment step S186). On the other hand, if the number of cells detached within the irradiated area is equal to or greater than the preset detachment rate (Yes in step S196), the scanning mechanism 15 moves to the next irradiation position (step S197). Then, when the ultrasonic beam 13 is irradiated over the entire cell 1 to detach the cell 1 from the culture vessel 2, the process of detaching the cell 1 for one cell 1 is completed.

[0127] Thereafter, if other cells 1 are adhered within one culture vessel 2, or if cells 1 are cultured in other wells of a multi-well plate, the cell detachment device 10 moves the transducer 11 to the next position, that is, to another cell 1 or another well in one culture vessel 2, by the scanning mechanism 15. Then, the cell detachment device 10 performs the series of steps S191 to S197 described above, and can detach a cell 1 of any shape from the cell adhesion surface of the culture vessel 2.

[0128] As described above, the cell detachment device 10 and cell detachment method according to this embodiment can irradiate the cells 1 being cultured with ultrasonic beams 13, thereby efficiently detaching the cells 1 in any shape from the culture vessel 2 in a non-contact manner. In particular, by detecting the detached state of the cells 1, the number of times that the ultrasonic beams 13 are irradiated to the cells 1 can be reduced, thereby further reducing damage to the cells 1 being detached.

[0129] An exemplary embodiment of this embodiment will be described below. [Aspect 1] A cell detachment device for detaching cells from a cell adhesion surface of a culture vessel, a plurality of transducers for irradiating the cells with ultrasonic beams through the culture vessel; An element driving unit that applies a voltage to a piezoelectric element of the vibrator; a scanning mechanism that moves the position of the transducer relative to the culture vessel; a control unit for controlling whether or not the ultrasonic beam is output, the beam output intensity, and the position of the transducer; having The transducers are arranged along an arbitrary array axis direction, The scanning mechanism scans the transducer along a scanning axis direction different from the array axis direction. A cell detachment device characterized by: [Aspect 2] The cell detachment device according to aspect 1, wherein the ultrasonic beam emitted by the transducer has a flat beam shape, and the long axis of the flat beam is arranged approximately perpendicular to the scanning axis direction. [Aspect 3] 2. The cell peeling device according to embodiment 1, further comprising a switching unit that switches the piezoelectric element to be driven. [Aspect 4] 2. The cell detachment device according to claim 1, wherein the transducer includes a beam variable mechanism for focusing the ultrasonic beam. [Aspect 5] The beam variable mechanism includes one or more acoustic lenses that focus the ultrasonic beam; The cell dissociation device according to aspect 4, wherein the control unit determines output conditions of the ultrasonic beam including focusing conditions of the ultrasonic beam by the acoustic lens. [Aspect 6] The beam variable mechanism includes a Fresnel lens that changes the focus of the ultrasonic beam, The cell detachment device according to aspect 4, wherein the control unit determines output conditions of the ultrasonic beam including a voltage to be applied by modulating a frequency to the piezoelectric element. [Aspect 7] a database having output conditions of the beam output of the ultrasonic beam according to at least one of the type of the cell and the type of the culture vessel; 2. The cell detachment apparatus according to claim 1, wherein the control unit sets output conditions of the ultrasonic beam by referring to the database. [Aspect 8] The cell detachment device according to aspect 7, wherein the control unit includes an external connection unit, and information in the database is updated with external information via the external connection unit. [Aspect 9] A detection unit that detects the detached state of the cells, The cell detachment device according to aspect 1, wherein the control unit determines the output of the ultrasonic beam and a scanning path for moving the transducer based on detachment state information output by the detection unit. [Aspect 10] The cell detachment device described in aspect 9, characterized in that the edge of the cell group detected by the detection unit is detached by irradiating the ultrasonic beam, and then the ultrasonic beam is irradiated to the boundary between the detached area where the cells have been detached and the adhesive area. [Aspect 11] The cell detachment device according to embodiment 1, further comprising a display unit that displays at least one of an actual image of the cell, the detached area, the shape of the cell, the mode of the ultrasonic beam, the scanning path, and the irradiation position. [Aspect 12] 2. The cell detachment device according to aspect 1, wherein a fluid is interposed between the culture vessel and the transducer. [Aspect 13] 13. The cell detachment device according to claim 12, wherein the fluid is water. [Aspect 14] 2. The cell detachment device according to claim 1, wherein the element driving unit outputs a burst wave of a wave packet having a fundamental frequency f0, the fundamental frequency f0 being equal to or higher than 150 kHz and equal to or lower than 50 MHz. [Aspect 15] 2. The cell detachment device according to claim 1, wherein the transducer emits a converging ultrasonic beam. [Aspect 16] The cell detachment device described in aspect 1, characterized in that the culture vessel is placed in a tank containing a fluid, the space between the culture vessel and the transducer is filled with the fluid, and the gap length between the culture vessel and the transducer is less than or equal to twice the wavelength of the ultrasonic beam in the fluid. [Aspect 17] 2. The cell detachment device according to aspect 1, wherein the array axis direction is substantially perpendicular to the scanning axis direction. [Aspect 18] The cell detachment device described in aspect 2, characterized in that adjacent transducers are positioned with a shift from each other in the direction of the short axis of the flat beam shape, and are positioned so that the amount of overlap between the positions of adjacent transducers is positive in the direction of the long axis. [Aspect 19] 3. The cell detachment device according to claim 2, wherein the width of the short axis of the flat shape is 5 mm or less. [Aspect 20] A cell detachment method for detaching cells from a cell adhesion surface of a culture vessel, A selection step of selecting a shape of a detachment area to be detached from the cell; a scanning path calculation step of calculating a scanning path of a transducer corresponding to the shape selected in the selection step; a moving step of moving the transducer to a designated position calculated and determined in the scanning path calculation step; a detachment step of applying a voltage to the transducer, generating ultrasonic waves from the transducer, and irradiating an ultrasonic beam to detach the cells from the cell adhesion surface; having The transducers are arranged along an arbitrary array axis direction, The moving step causes the transducer to scan along a scanning axis direction different from the array axis direction by a scanning mechanism that moves a position of the transducer relative to the culture vessel. A method for cell detachment comprising the steps of: [Aspect 21] A cell detachment method for detaching cells from a cell adhesion surface of a culture vessel, A selection step of selecting a shape of a detachment area to be detached from the cell; a scanning path calculation step of calculating a scanning path of a transducer corresponding to the shape selected in the selection step; a moving step of moving the transducer to a designated position calculated and determined in the scanning path calculation step; a detachment step of applying a voltage to the transducer, generating ultrasonic waves from the transducer, and irradiating an ultrasonic beam to detach the cells from the cell adhesion surface; a detection step of detecting a detached state of the cells at the location irradiated in the detachment step; an adjustment step of adjusting an output condition and an irradiation position of an ultrasonic beam for detaching the undetached cells detected in the detection step; having The transducers are arranged along an arbitrary array axis direction, The moving step causes the transducer to scan along a scanning axis direction different from the array axis direction by a scanning mechanism that moves a position of the transducer relative to the culture vessel. A method for cell detachment comprising the steps of: [Aspect 22] A cell detachment method described in aspect 20 or aspect 21, characterized in that the beam shape of the ultrasonic beam emitted by the transducer is flat, and the long axis of the flat shape is arranged approximately perpendicular to the scanning axis direction.

[0130] Although the cell detachment device and the cell detachment method according to the present invention have been described in detail by the embodiments, the present invention is not limited to the above-mentioned embodiments and includes various modified examples. For example, the above-mentioned embodiments have been described in detail to easily explain the present invention, and the present invention is not necessarily limited to those having all the described configurations. In addition, it is possible to replace a part of the configuration of one embodiment with the configuration of another embodiment, and it is also possible to add the configuration of another embodiment to the configuration of one embodiment. In addition, it is possible to add, delete, or replace a part of the configuration of each embodiment with another configuration. These embodiments and their modifications are included in the scope and gist of the invention, and are included in the scope of the invention and its equivalents described in the claims. [Explanation of symbols]

[0131] 1 cell 1-1 Cells in the irradiated area 1-2 Cells at the detachment boundary 2 Culture container 3 Culture solution 4 fluid 5 tanks 10 Cell detachment device 11 Transducer 12 Ultrasound probe 13 Ultrasonic beam 14 Element drive unit 15 Scanning mechanism 16 Control section 17 Signal Line 31 Switching section 41 Beam variable mechanism 51 Detection unit 52 Database 53 Beam control section 54 Beam condition determination unit 55 Scanning path calculation unit 56 Input section 57 Display section 58 Recognition part 59 Planning Department 310 Beam Shape 321 Long axis 322 Short axis 411 Piezoelectric element 412 Ground electrode 413 Signal Electrode 415 Protective film 511 Peeling Area 512 Non-peeling area

Claims

1. A cell detachment device for detaching cells from a cell adhesion surface of a culture vessel, a plurality of transducers for irradiating the cells with ultrasonic beams through the culture vessel; An element driving unit that applies a voltage to a piezoelectric element of the vibrator; a scanning mechanism that moves the position of the transducer relative to the culture vessel; a control unit for controlling whether or not the ultrasonic beam is output, the beam output intensity, and the position of the transducer; having The transducers are arranged along an arbitrary array axis direction, The scanning mechanism scans the transducer along a scanning axis direction different from the array axis direction. A cell detachment device characterized by:

2. 2. The cell detachment device according to claim 1, wherein the ultrasonic beam emitted by the transducer has a flat beam shape, and the major axis of the flat beam is disposed approximately perpendicular to the scanning axis direction.

3. 2. The cell detachment device according to claim 1, further comprising a switching unit for switching the piezoelectric element to be driven.

4. The cell detachment device according to claim 1 , wherein the transducer includes a beam variable mechanism for focusing the ultrasonic beam.

5. a database having output conditions of the beam output of the ultrasonic beam according to at least one of the type of the cell and the type of the culture vessel; The cell detachment apparatus according to claim 1 , wherein the control unit sets output conditions of the ultrasonic beam by referring to the database.

6. The cell detachment apparatus according to claim 5 , wherein the control unit includes an external connection unit, and information in the database is updated with external information via the external connection unit.

7. A detection unit that detects the detached state of the cells, The cell detachment device according to claim 1 , wherein the control unit determines the output of the ultrasonic beam and a scanning path for moving the transducer based on detachment state information output by the detection unit.

8. 2. The cell detachment device according to claim 1, wherein a fluid is interposed between the culture vessel and the transducer.

9. 2. The cell detachment device according to claim 1, wherein the element driving unit outputs a burst wave of a wave packet having a fundamental frequency f0, the fundamental frequency f0 being 150 kHz or more and 50 MHz or less.

10. The cell detachment device according to claim 1, characterized in that the culture vessel is placed in a tank containing a fluid, the space between the culture vessel and the transducer is filled with the fluid, and a gap length between the culture vessel and the transducer is less than or equal to twice the wavelength of the ultrasonic beam in the fluid.

11. The cell detachment device according to claim 2, characterized in that adjacent transducers are arranged so as to be offset from each other in the direction of the short axis of the flat beam shape, and are arranged so that the amount of overlap between the positions of adjacent transducers is positive in the direction of the long axis.

12. A cell detachment method for detaching cells from a cell adhesion surface of a culture vessel, A selection step of selecting a shape of a detachment area to be detached from the cell; a scanning path calculation step of calculating a scanning path of a transducer corresponding to the shape selected in the selection step; a moving step of moving the transducer to a designated position calculated and determined in the scanning path calculation step; a detachment step of applying a voltage to the transducer, generating ultrasonic waves from the transducer, and irradiating an ultrasonic beam to detach the cells from the cell adhesion surface; having The transducers are arranged along an arbitrary array axis direction, The moving step causes the transducer to scan along a scanning axis direction different from the array axis direction by a scanning mechanism that moves a position of the transducer relative to the culture vessel. A method for cell detachment comprising the steps of:

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