Cell separation device and method for controlling cell separation device

Abstract

To provide a cell separation device enabling appropriate control and efficient cell separation regardless of the combination with a culture substrate.SOLUTION: A signal output unit that outputs an AC signal, a vibration member that vibrates according to a frequency of the AC signal, a vibration transmission member that transmits vibration of the vibration member to the base material, an amplitude detection unit that detects an amplitude of vibration of the vibration transmission member, a resonance information acquisition unit that acquires resonance information, and a drive control unit that controls output of the AC signal based on the amplitude detected by the amplitude detection unit and the resonance information, wherein the resonance information includes resonance point information that is information related to a resonance amplitude peak of the vibration transmission member, the drive controller controls the output of the AC signal within a predetermined range between the frequency corresponding to a peak top and the frequency corresponding to one of a peak start and a peak end of the one resonance amplitude peak.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a technique for detaching cells from a culture substrate. [Background technology]

[0002] In the medical field, cells are sometimes cultured on culture substrates (hereinafter simply referred to as substrates) such as culture plates and dishes for treatment and research and development. After culture, the cells may adhere to the culture substrate. Therefore, in order to obtain sample cells after culture, it is necessary to detach the cells from the culture substrate used for culture.

[0003] There are two methods for detaching cells from a culture substrate: one is to add enzymes or chemicals that act on the cell membrane, and the other is to apply vibrational energy to the cells by applying acoustic radiation pressure.

[0004] Detachment methods using enzymes or chemicals that act on cell membranes can result in reduced cell proliferation and activity due to the dissolution of cellular proteins. For this reason, methods that apply vibrational energy to cells by applying acoustic radiation pressure to detach them from the culture substrate are preferred, but these tend to be costly and technically difficult. Specifically, for example, the optimal vibration amplitude for cell detachment methods using acoustic radiation pressure is determined by the trade-off between cell detachment rate and cell viability. Therefore, it is necessary to adjust the vibration amplitude to a stable level each time the cell detachment procedure is performed.

[0005] Patent Document 1 discloses a technique for detaching cells held in a container by irradiating the outer surface of the container in which the cells are cultured with ultrasonic waves in a frequency sweep manner. Patent Document 2 discloses a technology in which a frequency sweep operation is performed during a process in which ultrasonic waves are applied to a substrate to detach cells on the substrate, and the driving frequency is set to a newly identified, more favorable frequency. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Publication No. 2019-30260 [Patent Document 2] Special Publication No. 2023-527574 Summary of the Invention [Problem to be solved by the invention]

[0007] The technology described in Patent Document 1 is a cell detachment method that irradiates ultrasonic waves by sweeping the frequency, and while it can reliably operate at the natural frequency, it does not operate continuously at the natural frequency, which has the problem of lengthening the time required for the cell detachment process. In the technique described in Patent Document 2, there were cases where appropriate control could not be performed depending on the combination of the vibration generator and the culture substrate.

[0008] Therefore, an object of the present invention is to provide a cell detachment device that enables appropriate control and efficient cell detachment, regardless of the combination with the culture substrate. [Means for solving the problem]

[0009] A cell detachment device according to one aspect of the present invention is a cell detachment device for detaching cells on a substrate by applying vibration, the cell detachment device comprising: a signal output unit that outputs an AC signal for generating the vibration; a vibrating member that vibrates in response to the frequency of the AC signal; a vibration transmission member that vibrates in response to the vibration of the vibration member and transmits the vibration of the vibration member to the base material; an amplitude detection unit that detects the amplitude of vibration of the vibration transmission member; a resonance information acquiring unit that acquires resonance information that is information regarding a correspondence relationship between the frequency and the amplitude of the vibration transmitting member; a drive control unit that controls output of the AC signal from the signal output unit based on the amplitude detected by the amplitude detection unit and the resonance information; and the resonance information includes resonance point information that is information about a resonance amplitude peak of the vibration transmitting member, the drive control unit controls the output of the AC signal within a predetermined range between the frequency corresponding to a peak top of one of the resonance amplitude peaks and the frequency corresponding to either a peak start or a peak end. It is characterized by:

[0010] Furthermore, a method for controlling a cell detachment device according to another aspect of the present invention is a method for controlling a cell detachment device for detaching cells from a substrate by applying vibration to the cells, the method comprising: the cell detachment device includes a vibration member that vibrates in response to a frequency of an AC signal, and a vibration transmission member that vibrates in response to the vibration of the vibration member and transmits the vibration of the vibration member to the substrate; a signal output step of outputting the AC signal; an amplitude detection step of detecting the amplitude of vibration of the vibration transmission member; a resonance information acquisition step of acquiring resonance information which is information regarding a correspondence relationship between the frequency and the amplitude of the vibration transmitting member; a drive control step of controlling output of the AC signal based on the amplitude detected in the amplitude detection step and the resonance information; and the resonance information includes resonance point information that is information about a resonance amplitude peak of the vibration transmitting member, the drive control step includes controlling the output of the AC signal within a predetermined range between the frequency corresponding to a peak top of one of the resonance amplitude peaks and the frequency corresponding to either a peak start or a peak end. It is characterized by: [Effects of the Invention]

[0011] According to the present invention, it is possible to provide a cell detachment device that enables appropriate control and efficient cell detachment regardless of the combination with the culture substrate. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is a functional block diagram of a cell detachment device according to a first embodiment of the present invention. FIG. [Figure 2] 1 is a diagram showing a system configuration of a cell detachment device according to a first embodiment of the present invention. [Figure 3] FIG. 2 is an exploded view showing the configuration of a vibration unit of the cell detachment device according to the first embodiment of the present invention. [Figure 4] 10(a) and 10(b) are graphs showing an example of the relationship between the frequency of an AC signal and the amplitude of a vibration transmitting member. [Figure 5] 10 is a flowchart illustrating an example of a method for acquiring resonance information. [Figure 6] 4 is a flowchart showing an example of control of the cell detachment device according to the first embodiment of the present invention. [Figure 7] 10 is a graph showing an example of the relationship between the frequency of an AC signal and the amplitude of a vibration transmitting member. [Figure 8] 10 is a flowchart showing an example of control of a cell detachment device according to a second embodiment of the present invention. [Figure 9] (a) is a graph showing the change in the natural frequency corresponding to one resonance point of the vibrator member over driving time, and (b) is a graph showing the relationship between the natural frequency corresponding to one resonance point and the surface temperature of the vibrator member. DETAILED DESCRIPTION OF THE INVENTION

[0013] Although the technology described in Patent Document 1 has the advantage of being able to reliably perform detachment at the natural frequency, in a cell detachment method using acoustic radiation pressure, it is desirable to perform the detachment process continuously at a constant frequency in order to complete the cell detachment process in a short time. Furthermore, a method for detaching cells from a substrate requires control that can obtain a wide range of vibration amplitude to accommodate various cell types, but in a detachment method using acoustic radiation pressure, it is difficult to obtain a wide range of vibration amplitude using voltage control alone, so frequency control is required.

[0014] In contrast, the technology described in Patent Document 2 performs a frequency sweep operation during the cell detachment process, thereby performing cell detachment at a more appropriate frequency that is identified, making it possible to perform continuous detachment at the identified preferred frequency. However, according to the investigations of the present inventors, the technology described in Patent Document 2 sometimes fails to perform appropriate control depending on the combination of the vibration generator and the culture substrate.

[0015] The method described in Patent Document 2 is based on the premise that control is performed for one natural frequency. However, depending on the combination of the vibration generator with the culture substrate, the vibration generator may have multiple natural frequencies. Therefore, it is considered that the technology described in Patent Document 2 may not be able to perform appropriate control when the vibration generator's natural frequency is combined with the culture substrate resulting in multiple natural frequencies. The present inventors have conducted studies to solve the above problems.

[0016] To control the frequency of a vibration generator with multiple different natural frequencies, it is necessary to obtain information on two natural frequencies: a frequency corresponding to at least one natural frequency, and a frequency at which the vibration amplitude between the natural frequency and the adjacent natural frequency is minimized.A control system is then required that can obtain the desired vibration amplitude based on the information on the two natural frequencies.

[0017] For this purpose, the above-mentioned two pieces of natural frequency information must be obtained before the start of the peeling process, and it is preferable to perform frequency control in which the information is also obtained during the peeling process and updated as appropriate.

[0018] The present inventors have further studied the present invention and have come up with the present invention. Hereinafter, each configuration of the present invention will be described in detail using exemplary embodiments of the present invention with reference to the drawings. In the drawings, similar or corresponding elements are designated by the same reference numerals, and their description may be omitted or simplified.

[0019] First Embodiment Fig. 1 is a functional block diagram of a cell detachment device according to a first embodiment of the present invention. Fig. 2 is a diagram showing the overall system configuration of a cell detachment device according to a first embodiment of the present invention. Fig. 3 is a schematic diagram of a vibration unit of a cell detachment device according to a first embodiment of the present invention, shown as an exploded view for ease of explanation.

[0020] The cell detachment device 10 according to this embodiment is a cell detachment device for detaching cells from a substrate 201 by applying vibrations to the cells on the substrate 201, and as shown in FIGS. 1 and 2, comprises a vibration applying unit 110, a vibration control unit 120, and a host control unit 130. The vibration applying unit 110 has a vibrating member 111, a vibration transmission member 112, and an amplitude detection unit 113. The vibration control unit 120 has a signal output unit 121, a drive control unit 122, an amplitude information acquisition unit 123, a resonance information acquisition unit 124, and a resonance information storage unit 125.

[0021] The vibration control unit 120 is a vibrator controller driver that outputs a signal for driving the vibrating member 111 of the vibration unit 110, and the vibration control unit 120 is operated by a host control unit 130 that has a GUI (Graphical User Interface) for human operation. The vibration control unit 120 and the host control unit 130 can be configured by devices such as a PC (Personal Computer) and a PLC (Programmable Logic Controller).

[0022] The signal output unit 121 outputs an AC signal for causing vibration in the vibration unit 110, and the vibrating member 111 vibrates according to the frequency of the AC signal output from the signal output unit 121. The AC signal output from the signal output unit 121 is a sine wave signal of a desired frequency. Specifically, for example, a Langevin vibrator can be used as the vibrating member 111, but the vibrating member 111 is not limited to this, and other types of vibrators using a resonance phenomenon may also be used.

[0023] The vibration transmission member 112 vibrates in response to the vibrations of the vibrating member 111, and transmits the vibrations of the vibrating member 111 to the base material 201. Specifically, for example, silicone rubber or the like can be used as the vibration transmission member 112. The vibration transmission member 112 may be made of any material that can transmit vibrations, and may be a liquid such as pure water, for example, but is not limited to such a material. When the vibration transmission member 112 is a liquid, a dike structure may be provided around the vibrating member 111 to prevent the liquid vibration transmission member 112 from flowing down.

[0024] For example, a culture medium and cells are provided on the substrate 201, and the cultured cells are adhered to the bottom surface 201a of the substrate 201. A weight 202 is placed on the top surface of the substrate 201.

[0025] The vibrating member 111 is disposed on the device housing (not shown) via the buffer member 203. That is, the lower surface of the vibrating member 111 is supported by the buffer member 203. Specifically, the buffer member 203 may be made of felt, for example.

[0026] The amplitude detection unit 113 detects the amplitude of vibration of the vibration transmission member 112. Specific examples of the amplitude detection unit 113 include a sensor that outputs a voltage according to the vibration amplitude using a piezoelectric member, a laser displacement meter, and the like.

[0027] The amplitude information acquisition unit 123 acquires information about the amplitude of the vibration transmitting member 112 detected by the amplitude detection unit 113 .

[0028] On the other hand, the resonance information acquisition unit 124 acquires resonance information, which is information about the correspondence between the frequency of the AC signal for generating vibration and the amplitude of the vibration transmission member 112 .

[0029] 4(a) and (b) show examples of graphs representing resonance information, with the horizontal axis plotting the frequency of the AC signal and the vertical axis plotting the amplitude of the vibration transmission member 112. Fig. 4(b) is an enlarged view of the portion indicated by the dashed line in Fig. 4(a).

[0030] 4(a), the amplitude of the vibration transmission member 112 may exhibit multiple resonance amplitude peaks when the frequency of the AC signal is changed, and each resonance amplitude peak corresponds to a multiple natural frequency of the vibration member 111. The resonance information includes resonance point information, which is information related to the resonance amplitude peaks of the vibration transmission member 112.

[0031] The resonance information is specified in advance and stored in the resonance information storage unit 125, and is acquired from the resonance information storage unit 125 by the resonance information acquisition unit 124 when the cell detachment device 10 is in operation.

[0032] Alternatively, the resonance information may be identified by the cell detachment device 10 immediately before detaching the cells and stored in the resonance information storage unit 125. Specifically, the drive control unit 122 is configured to be able to control the output of the AC signal by the signal output unit 121 so as to sweep the frequency within a predetermined range, and before detaching the cells, the signal output unit 121 sweeps the frequency and outputs the AC signal.

[0033] An example of a specific operation for identifying resonance information by the cell detachment device 10 is shown in Fig. 5. In the example shown in Fig. 5, a case where the frequency is swept from a high frequency to a low frequency is described, but the frequency sweeping method is not limited to this, and for example, the frequency may be swept from a low frequency to a high frequency.

[0034] First, in step S101, the drive control unit 122 determines sweep conditions including a frequency range to be swept and a step frequency. The sweep conditions may be input by a user from, for example, the upper control unit 130. The frequency range to be swept is set so as to cover the range of the natural frequency of the vibrating member 111.

[0035] Next, in step S102, the signal output unit 121 starts outputting an AC signal from a predetermined start frequency F0 based on the sweep conditions. Accordingly, the vibrating member 111 vibrates in response to the AC signal, and the vibration transmission member 112 vibrates in response to the vibration of the vibrating member 111. The amplitude of the vibration of the vibration transmission member 112 is then detected by the amplitude detection unit 113.

[0036] Subsequently, in step S103 , the amplitude information acquisition unit 123 acquires information about the amplitude of the vibration transmitting member 112 detected by the amplitude detection unit 113 .

[0037] Next, in step S104, the drive control unit 122 transmits the next command to the signal output unit 121 depending on whether the amplitude of the vibration transmission member 112 acquired by the amplitude information acquisition unit 123 is larger or smaller than a reference value. Here, the initial value of the reference value is set to 0.

[0038] If the amplitude of the vibration transmission member 112 acquired by the amplitude information acquisition unit 123 is greater than the reference value, the drive control unit 122 sends a command to the signal output unit 121 to output an AC signal with a frequency that is lower by the step frequency. At the same time, the drive control unit 122 updates the reference value to the amplitude value acquired by the amplitude information acquisition unit 123. The process then returns to step S102, where the signal output unit 121 outputs an AC signal based on the received command. In step S103, the amplitude information acquisition unit 123 acquires information about the amplitude of the vibration transmission member 112 that corresponds to the changed frequency. In step S104, the drive control unit 122 compares the amplitude of the vibration transmission member 112 newly acquired by the amplitude information acquisition unit 123 with the updated reference value. The above operation is repeated until the amplitude acquired by the amplitude information acquisition unit 123 becomes smaller than the reference value.

[0039] If the amplitude acquired by the amplitude information acquisition unit 123 becomes smaller than the reference value, the drive control unit 122 updates the start frequency F0 to the frequency at that time in step S105.

[0040] Subsequently, in step S106, the signal output unit 121 outputs an AC signal with the updated start frequency F0.

[0041] Thereafter, in step S107 , the amplitude information acquisition unit 123 acquires information about the amplitude of the vibration transmitting member 112 detected by the amplitude detection unit 113 .

[0042] Next, in step S108, the drive control unit 122 transmits the next command to the signal output unit 121 depending on whether the amplitude of the vibration transmission member 112 acquired by the amplitude information acquisition unit 123 is large or small relative to a reference value.

[0043] If the amplitude of the vibration transmission member 112 acquired by the amplitude information acquisition unit 123 is smaller than the reference value, the drive control unit 122 checks whether the termination condition is met in step S109. If the termination condition is not met, the drive control unit 122 sends a command to the signal output unit 121 to output an AC signal with a frequency lower by the step frequency. At the same time, the drive control unit 122 updates the reference value to the amplitude value acquired by the amplitude information acquisition unit 123. Then, the process returns to step S106, and the signal output unit 121 outputs an AC signal based on the received command.

[0044] If the amplitude acquired by the amplitude information acquisition unit 123 becomes larger than the reference value, the drive control unit 122 updates the start frequency F0 to the frequency at that time in step S110. Next, the process returns to step S102, where the signal output unit 121 outputs an AC signal with the updated start frequency F0.

[0045] If the termination condition is met in step S109, in step S111 the drive control unit 122 commands the signal output unit 121 to terminate the frequency sweep. The termination condition can be set arbitrarily, but for example, the termination condition can be set to be that the flow of steps S106 to S109 has been repeated a predetermined number of times.

[0046] The resonance information acquiring unit 124 can be configured to identify and acquire resonance point information in the process of sweeping a predetermined frequency range. Specifically, the resonance information acquiring unit 124 identifies and acquires, as resonance point information, at least one of the peak top, peak start, and peak end of the resonance amplitude peak based on the amplitude detected by the amplitude detecting unit 113.

[0047] 4(a), the peak tops of the resonance amplitude peaks are designated as resonance points A, B, and C in order from highest frequency. Since it is preferable to control the frequency based on resonance point B where the maximum amplitude is obtained, the resonance information acquisition unit 124 identifies the frequency and amplitude at resonance point B as resonance point information 2. Furthermore, the resonance information acquisition unit 124 identifies the frequency and amplitude at point A', which corresponds to the minimum value of the amplitude between resonance point A and resonance point B, as resonance point information 1.

[0048] Based on the amplitude and resonance information detected by the amplitude detection unit 113, the drive control unit 122 controls the output of the AC signal from the signal output unit 121 so that the vibration transmission member 112 has a target amplitude. Here, the drive control unit 122 controls the output of the AC signal within a predetermined range from the frequency corresponding to the peak top of one resonance amplitude peak to the frequency corresponding to either the peak start or peak end. The drive control unit 122 controls at least one of the frequency of the AC signal and the applied voltage for outputting the AC signal.

[0049] When the resonance point information includes information on a main peak, which is two adjacent resonance amplitude peaks, and a sub-peak, which has a smaller amplitude at its peak top than the main peak, the drive control unit 122 preferably controls the output of the AC signal as follows: That is, the drive control unit 122 preferably controls the output of the AC signal within a predetermined range from the frequency corresponding to the peak top of the main peak to the frequency corresponding to the minimum value of the amplitude between the two resonance amplitude peaks.

[0050] In the example shown in Figures 4(a) and (b), the resonance amplitude peak including resonance point B, which has the largest amplitude at the peak top, can be considered as the main peak, and the resonance amplitude peak including resonance point A, which is adjacent to the resonance amplitude peak including resonance point B, can be considered as the sub-peak. The frequency between resonance point information 1 and resonance point information 2 can be set as the control range. However, driving at a frequency at the peak top of the resonance amplitude peak makes the vibration amplitude unstable, so it is preferable that the drive control unit 122 control the output of the AC signal within a frequency range excluding the frequency at the peak top. For example, in the example shown in Figure 4(b), it is preferable to set the drive range to a frequency slightly higher than that of resonance point information 2, and specifically, for example, (resonance point information 2) + 100 Hz is set as the lower limit of the drive frequency.

[0051] Because the range of frequencies controlled by the drive control unit 122 is limited, the drive control unit 122 performs control, including adjusting the applied voltage as appropriate, so as not to exceed the limits of that range. Specifically, for example, in the example shown in FIG. 4(b), consider a case where the control range is from resonance point information 1 to (resonance point information 2) + 100 Hz. In this case, if the amplitude is larger than the target amplitude even when the drive frequency is set to resonance point information 1, the drive control unit 122 performs control to reduce the applied voltage for outputting the AC signal from the signal output unit 121. On the other hand, if the amplitude is smaller than the target amplitude even when the drive frequency is set to (resonance point information 2) + 100 Hz, the drive control unit 122 performs control to increase the applied voltage for outputting the AC signal from the signal output unit 121.

[0052] If the peak start or peak end, which is the reference when the drive control unit 122 controls the frequency, is not an inflection point between adjacent resonance amplitude peaks, for example, if there is one resonance amplitude peak, the peak start or peak end may be determined as follows: That is, the point at which the amount of change in the amplitude of the vibration transmitting member 112 in response to a frequency fluctuation becomes equal to or less than a predetermined threshold value can be determined as the peak start or peak end.

[0053] Generally, the resolution for controlling the frequency of an AC signal is lower than the resolution for controlling the applied voltage for outputting the AC signal, and the relationship between frequency and amplitude at the resonance amplitude peak exhibits a steep behavior. Therefore, it is preferable to adjust large amplitudes by controlling the frequency and to adjust small amplitudes by controlling the applied voltage.

[0054] Specifically, it is preferable that the drive control unit 122 performs the following control based on the difference between the amplitude of the vibration transmission member 112 detected by the amplitude detection unit 113 and the target amplitude. That is, if the difference is larger than the amount of change in the amplitude of the vibration transmission member 112 predicted from the resonance information when the frequency of the AC signal output from the signal output unit 121 is changed by the minimum amount, the drive control unit 122 changes the frequency of the AC signal. On the other hand, if the difference is smaller than the amount of change in the amplitude of the vibration transmission member 112 predicted from the resonance information when the frequency of the AC signal output from the signal output unit 121 is changed by the minimum amount, the drive control unit 122 changes the applied voltage for outputting the AC signal.

[0055] Next, a control method for the cell detachment device according to this embodiment will be described. The control method for the cell detachment device according to this embodiment can be realized by the components of the cell detachment device 10 described above.

[0056] FIG. 6 is a diagram showing an example of a control flow of the cell detachment device 10 in the cell detachment step. First, in a resonance information acquisition step of step S201, the resonance information acquisition unit 124 acquires resonance information.

[0057] Next, in the drive condition determination process of step S202, drive conditions including the target amplitude, control frequency range, drive frequency, applied voltage (power supply voltage), and termination condition are determined. For example, the control frequency range is determined based on the resonance point information 1 and 2 in the example described above with reference to FIG. 4. The termination condition can be, for example, a predetermined drive time. The drive frequency and applied voltage can be determined by estimating the combination of drive frequency and applied voltage that will obtain the target amplitude based on the resonance information.

[0058] The driving conditions may be input by the user from the host control unit 130, or may be automatically determined by the cell detachment device 10 from the resonance information acquired by the resonance information acquisition unit 124 and the target amplitude for detachment. In this case, the target amplitude for detachment may be input by the user from the host control unit 130, or may be automatically determined by the cell detachment device 10 from information including the cell type input by the user from the host control unit 130.

[0059] Next, in a signal output step S203, the signal output unit 121 outputs an AC signal based on the driving conditions.

[0060] Subsequently, in an amplitude detection step of step S204, the amplitude of the vibration of the vibration transmitting member 112 is detected by the amplitude detection unit 113.

[0061] Next, in the termination determination step of step S205, the drive control unit 122 determines whether or not the termination condition is satisfied. If it is determined that the termination condition is not satisfied, the process proceeds to the drive control step of step S206.

[0062] In the drive control process of step S206, the output of the AC signal from the signal output unit 121 is controlled based on the amplitude of the vibration transmission member 112 detected in the amplitude detection process and the resonance information acquired in the resonance information acquisition process so that the vibration transmission member 112 has a target amplitude. In this embodiment, the drive control process includes controlling the output of the AC signal within a predetermined range between a frequency corresponding to the peak top of one resonance amplitude peak and a frequency corresponding to either the peak start or the peak end.

[0063] In the drive control process, as described above, it is preferable that the drive control unit 122 controls to change the frequency of the AC signal or the applied voltage for outputting the AC signal based on the difference between the amplitude of the vibration transmission member 112 detected by the amplitude detection unit 113 and the target amplitude.

[0064] If it is determined in the termination determination process of step S205 that the termination conditions are met, the driving of the cell detachment device 10 is terminated in the termination process of step S207. The termination conditions for the detachment process may be set by setting the driving end time based on the results of a detachment experiment in advance, or the termination determination may be made based on images captured by a separate camera or the like.

[0065] According to this embodiment, it is possible to provide a cell detachment device that enables appropriate control and efficient cell detachment regardless of the combination with the culture substrate. Specifically, according to this embodiment, stable control is possible even when the combination of the substrate 201, the vibrating member 111, and the vibration transmitting member 112 has multiple resonance amplitude peaks.

[0066] 7 is an example of a graph showing the relationship between the frequency of an AC signal and the amplitude of the vibration transmission member 112 when three mutually different substrates 201 are combined with the vibration member 111 and the vibration transmission member 112. Resonance point information acquired when the substrate A is combined with the vibration member 111 and the vibration transmission member 112 is defined as A1 and A2, and the frequency control range for the substrate A is defined as A1 to A2+x (Hz). Here, x is a positive number that is determined as appropriate so that the frequency in the resonance point information A2 is not included in the frequency control range.

[0067] Consider the case where cell type 1 and cell type 2, which have different preferred detachment conditions, are to be detached. Here, cell type 1 is a cell type that has strong adhesion to the dish and requires a large vibration amplitude to detach the cells, and the vibration amplitude required for cell type 1 is set as target amplitude 1. Cell type 2 has weak adhesion to itself and, for example, when detaching the cells into a sheet, is a cell type that easily tears the cell sheet with strong vibration, and the vibration amplitude required for cell type 2 is set as target amplitude 2. In this case, it is necessary to obtain a range of amplitudes that includes target amplitude 1 and target amplitude 2 through control.

[0068] If frequency control of substrate B or substrate C is performed using resonance point information A2 for substrate A, cell type 1 will not be able to obtain the required vibration amplitude. Similarly, if frequency control of substrate B or substrate C is performed using resonance point information A1, cell type 2 will be subjected to excessive vibration amplitude. For this reason, frequency control must be performed based on resonance point information for the combination of each substrate 201, vibrating member 111, and vibration transmission member 112.

[0069] Furthermore, Figure 7 shows that target amplitude 1 required for detaching cell type 1 has two frequency solutions for each of substrates A to C. The same is true for target amplitude 2 required for detaching cell type 2. Therefore, stable control is difficult to achieve with conventional control methods for cell detachment devices that do not impose any particular restrictions on the frequency control range.

[0070] As described above, according to this embodiment, even in a vibration generator having multiple natural vibrations, it is possible to control the vibration amplitude when vibration is applied to a desired value, thereby achieving both an optimal cell detachment rate and an optimal cell survival rate.

[0071] In this embodiment, one limit value of the frequency range to be controlled is not necessarily limited to the frequency at the peak start or peak end of the resonance amplitude peak. That is, in the example shown in Fig. 4(b), for example, resonance point information 2 does not necessarily have to be frequency A' at the minimum amplitude, but may be a frequency that results in a vibration amplitude that is sufficiently lower than the vibration amplitude obtained by resonance point information 1. As long as the vibration amplitude value is sufficiently low, resonance point information 2 may be a fixed value based on the results of a previous measurement.

[0072] Second Embodiment FIG. 8 is a flowchart showing an example of control of the cell detachment device according to the second embodiment of the present invention.

[0073] The main differences between this embodiment and the first embodiment will be described below. In this embodiment, the drive control unit 122 is configured to be able to control the output of the AC signal from the signal output unit 121 so as to sweep the frequency at a predetermined timing during the operation of the cell detachment device 10. Furthermore, the resonance information acquisition unit 124 is configured to be able to update the resonance point information based on the results of the frequency sweep at the predetermined timing.

[0074] In controlling the cell detachment device 10 in this embodiment, the procedure from step S201 to step S204 shown in FIG. 8 is the same as that described in the first embodiment.

[0075] If the termination condition is not satisfied in the termination determination process of step S205, in this embodiment, the drive control unit 122 next determines whether a predetermined sweep operation condition is satisfied in the sweep determination process of step S301. The sweep operation condition is not particularly limited, and for example, the sweep operation may be performed every time a predetermined drive time (e.g., one minute) has elapsed. Furthermore, the sweep operation condition is not limited to the drive time described above. For example, the surface temperature of the vibrating member 111 may be measured separately, and the sweep operation may be performed based on the measured temperature change. Furthermore, the sweep operation may be performed when the desired amplitude of the vibration transmission member 112 can no longer be obtained within the control range of the drive frequency when an AC signal is output at a predetermined applied voltage.

[0076] If it is determined in the sweep determination step that the sweep operation conditions are not satisfied, the process proceeds to the drive control step of step S206. The operation in the drive control step is the same as that described in the first embodiment.

[0077] If it is determined in the sweep determination step that the sweep operation conditions are met, then in the frequency sweep step of step S302, the drive control unit 122 controls the output from the signal output unit 121 so as to perform a sweep within a predetermined frequency range.

[0078] Thereafter, the process returns to the resonance information acquisition step of step S201, where the resonance information acquisition unit 124 acquires and updates the resonance point information based on the results of the sweep operation.

[0079] According to this embodiment, the control conditions can be appropriately controlled at any time during the operation of the cell detachment device 10 to detach cells.

[0080] Specifically, for example, in a cell detachment device that detaches cells by applying vibration, the natural frequency of the vibrating member may fluctuate due to self-heating during the detachment process. FIG. 9 is a graph showing an example of fluctuations in the natural frequency of the vibrating member during the operation of the cell detachment device. FIG. 9(a) is a graph showing the change in the natural frequency corresponding to one resonance point of the vibrating member over drive time, and FIG. 9(b) is a graph showing the relationship between the natural frequency corresponding to one resonance point and the surface temperature of the vibrating member. Of the applied voltages 1 to 3 in FIG. 9, applied voltage 1 is the lowest applied voltage, and applied voltage 3 is the highest applied voltage. As shown in FIG. 9(a), the frequency of the resonance point may decrease with the drive time of the cell detachment device. Furthermore, as shown in FIG. 9(b), the frequency of the resonance point may decrease with an increase in the surface temperature of the vibrating member. Therefore, by controlling the cell detachment device according to this embodiment, cell detachment can always be performed with an appropriate vibration amplitude in accordance with fluctuations in the natural frequency during operation.

[0081] It should be noted that the above-described embodiments are merely illustrative examples of the present invention, and the technical scope of the present invention should not be construed as being limited by them. That is, the present invention can be embodied in various forms without departing from its technical concept or main features. For example, it should be understood that embodiments in which part of the configuration of any of the embodiments is added to or substituted for part of the configuration of another embodiment are also embodiments to which the present invention can be applied.

[0082] For example, the cell detachment device according to the present invention is not limited to the above-described configuration, and may be configured to generate vibrations in the ultrasonic band. Furthermore, the present invention may be realized by a cell detachment system comprising multiple devices connected by wire or wirelessly.

[0083] The disclosure according to the embodiments of the present invention includes the following configurations and methods. (Configuration 1) A cell detachment device for detaching cells on a substrate from the substrate by applying vibration, comprising: a signal output unit that outputs an AC signal for generating the vibration; a vibrating member that vibrates in response to the frequency of the AC signal; a vibration transmission member that vibrates in response to the vibration of the vibration member and transmits the vibration of the vibration member to the base material; an amplitude detection unit that detects the amplitude of vibration of the vibration transmission member; a resonance information acquiring unit that acquires resonance information that is information regarding a correspondence relationship between the frequency and the amplitude of the vibration transmitting member; a drive control unit that controls output of the AC signal from the signal output unit based on the amplitude detected by the amplitude detection unit and the resonance information; and the resonance information includes resonance point information that is information about a resonance amplitude peak of the vibration transmitting member, the drive control unit controls the output of the AC signal within a predetermined range between the frequency corresponding to a peak top of one of the resonance amplitude peaks and the frequency corresponding to either a peak start or a peak end. A cell detachment device characterized by: (Configuration 2) the resonance point information includes information about a main peak, which is two adjacent resonance amplitude peaks, and a sub-peak, the amplitude of which at the peak top is smaller than that of the main peak; The cell detachment device described in configuration 1, wherein the drive control unit controls the output of the AC signal within a predetermined range between the frequency corresponding to the peak top of the main peak and the frequency corresponding to the minimum value of the amplitude between the two resonant amplitude peaks. (Configuration 3) The cell detachment device according to configuration 1 or 2, wherein the drive control unit changes the frequency of the AC signal when the difference between the amplitude detected by the amplitude detection unit and a target amplitude is larger than the amount of change in the amplitude of the vibration transmission member predicted from the resonance information when the frequency is changed at a minimum, and changes the applied voltage for outputting the AC signal when the difference between the amplitude detected by the amplitude detection unit and a target amplitude is smaller than the amount of change in the amplitude of the vibration transmission member predicted from the resonance information when the frequency is changed at a minimum. (Configuration 4) the drive control unit is configured to be able to control the output of the AC signal so as to sweep the frequency within a predetermined range, The cell detachment device of configuration 3, wherein the resonance information acquisition unit is configured to be able to identify and acquire the peak top, peak start, and / or peak end of the resonance amplitude peak as the resonance point information based on the amplitude detected by the amplitude detection unit during the process of sweeping the frequency within the specified range. (Configuration 5) the drive control unit is configured to be able to control the output of the AC signal so as to sweep the frequency at a predetermined timing during the operation of the cell detachment device, 5. The cell detachment device according to claim 4, wherein the resonance information acquisition unit is configured to be able to update the resonance point information based on the result of sweeping the frequency at the predetermined timing. (Configuration 6) 6. The cell detachment device according to any one of configurations 1 to 5, wherein the drive control unit controls at least one of the frequency of the AC signal and an applied voltage for outputting the AC signal. (Configuration 7) 7. The cell detachment device according to any one of configurations 1 to 6, wherein the combination of the base material, the vibration member, and the vibration transmission member has a plurality of resonance amplitude peaks. (Configuration 8) 8. The cell detachment device according to any one of configurations 1 to 7, wherein the drive control section controls the output of the AC signal within the frequency range excluding the frequency at the peak top. (Method 1) 1. A method for controlling a cell detachment device for detaching cells from a substrate by applying vibration to the cells, the method comprising: the cell detachment device includes a vibration member that vibrates in response to a frequency of an AC signal, and a vibration transmission member that vibrates in response to the vibration of the vibration member and transmits the vibration of the vibration member to the substrate; a signal output step of outputting the AC signal; an amplitude detection step of detecting the amplitude of vibration of the vibration transmission member; a resonance information acquisition step of acquiring resonance information which is information regarding a correspondence relationship between the frequency and the amplitude of the vibration transmitting member; a drive control step of controlling output of the AC signal based on the amplitude detected in the amplitude detection step and the resonance information; and the resonance information includes resonance point information that is information about a resonance amplitude peak of the vibration transmitting member, the drive control step includes controlling the output of the AC signal within a predetermined range between the frequency corresponding to a peak top of one of the resonance amplitude peaks and the frequency corresponding to either a peak start or a peak end. A control method comprising: [Explanation of symbols]

[0084] 10 Cell detachment device 110 Vibration unit 111 Vibration member 112 Vibration transmission member 113 Amplitude detection unit 120 Vibration control unit 121 Signal output section 122 Drive control unit 123 Amplitude information acquisition section 124 Resonance information acquisition section 130 Upper control unit

Claims

1. A cell detachment device for detaching cells on a substrate from the substrate by applying vibration, comprising: a signal output unit that outputs an AC signal for generating the vibration; a vibrating member that vibrates in response to the frequency of the AC signal; a vibration transmission member that vibrates in response to the vibration of the vibration member and transmits the vibration of the vibration member to the base material; an amplitude detection unit that detects the amplitude of vibration of the vibration transmission member; a resonance information acquiring unit that acquires resonance information that is information regarding a correspondence relationship between the frequency and the amplitude of the vibration transmitting member; a drive control unit that controls output of the AC signal from the signal output unit based on the amplitude detected by the amplitude detection unit and the resonance information; and the resonance information includes resonance point information that is information about a resonance amplitude peak of the vibration transmitting member, the drive control unit controls the output of the AC signal within a predetermined range from the frequency corresponding to a peak top to the frequency corresponding to either a peak start or a peak end of one of the resonance amplitude peaks. A cell detachment device characterized by:

2. the resonance point information includes information about a main peak, which is two adjacent resonance amplitude peaks, and a sub-peak, the amplitude of which at the peak top is smaller than that of the main peak; The cell detachment device described in claim 1, wherein the drive control unit controls the output of the AC signal within a predetermined range between the frequency corresponding to the peak top of the main peak and the frequency corresponding to the minimum value of the amplitude between the two resonant amplitude peaks.

3. 2. The cell detachment device according to claim 1, wherein the drive control unit changes the frequency of the AC signal when a difference between the amplitude detected by the amplitude detection unit and a target amplitude is greater than an amount of change in the amplitude of the vibration transmission member predicted from the resonance information when the frequency is changed at a minimum, and changes the applied voltage for outputting the AC signal when a difference between the amplitude detected by the amplitude detection unit and a target amplitude is smaller than an amount of change in the amplitude of the vibration transmission member predicted from the resonance information when the frequency is changed at a minimum.

4. the drive control unit is configured to be able to control the output of the AC signal so as to sweep the frequency within a predetermined range, The cell detachment device of claim 3, wherein the resonance information acquisition unit is configured to be able to identify and acquire the peak top, peak start, and / or peak end of the resonance amplitude peak as the resonance point information based on the amplitude detected by the amplitude detection unit during the process of sweeping the frequency within the specified range.

5. the drive control unit is configured to be able to control the output of the AC signal so as to sweep the frequency at a predetermined timing during the operation of the cell detachment device, The cell detachment device according to claim 4 , wherein the resonance information acquisition unit is configured to be able to update the resonance point information based on a result of sweeping the frequency at the predetermined timing.

6. The cell detachment device according to claim 1 , wherein the drive control unit controls at least one of the frequency of the AC signal and an applied voltage for outputting the AC signal.

7. The cell detachment device according to claim 1 , wherein the combination of the substrate, the vibration member, and the vibration transmission member has a plurality of resonance amplitude peaks.

8. The cell detachment device according to claim 1 , wherein the drive control unit controls the output of the AC signal within the frequency range excluding the frequency at the peak top.

9. 1. A method for controlling a cell detachment device for detaching cells from a substrate by applying vibration to the cells, the method comprising: the cell detachment device includes a vibration member that vibrates in response to a frequency of an AC signal, and a vibration transmission member that vibrates in response to the vibration of the vibration member and transmits the vibration of the vibration member to the substrate; a signal output step of outputting the AC signal; an amplitude detection step of detecting the amplitude of vibration of the vibration transmission member; a resonance information acquisition step of acquiring resonance information which is information regarding a correspondence relationship between the frequency and the amplitude of the vibration transmitting member; a drive control step of controlling output of the AC signal based on the amplitude detected in the amplitude detection step and the resonance information; and the resonance information includes resonance point information that is information about a resonance amplitude peak of the vibration transmitting member, the drive control step includes controlling the output of the AC signal within a predetermined range between the frequency corresponding to a peak top of one of the resonance amplitude peaks and the frequency corresponding to either a peak start or a peak end of the one of the resonance amplitude peaks. A control method comprising:

Images