Cell observation system

Abstract

To provide a cell observation system capable of observing a cell while reducing blurring of an observation image of the cell.SOLUTION: Provided is an observation system including a cell separation device that separates cells provided on a culture surface of a culture vessel by applying vibration to the culture vessel, and an imaging device having an illumination unit that irradiates the cells with light and an imaging unit that images the cells, wherein the illumination unit periodically repeats light emission and light extinction, and a continuous light emission time of the light is shorter than a period of the vibration.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a cell observation system. [Background technology]

[0002] In the fields of regenerative medicine and drug discovery, methods for producing cell products by culturing various cells have been developed. In particular, when expanding adherent cells that adhere to a culture substrate, a passaging procedure is performed in which the cultured cells are detached from the substrate and seeded. Therefore, there is a need for a minimally invasive method for detaching cells that have adhered to the substrate. Meanwhile, a cell detachment method using ultrasound has been proposed. Patent Document 1 discloses a cell detachment device that detaches cells by applying vibrations to a culture vessel. The cell detachment device in Patent Document 1 captures images of the inside of a dish to generate image data. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2016 / 047368 Summary of the Invention [Problem to be solved by the invention]

[0004] When performing a cell detachment method in which cells are detached by applying vibration to a culture vessel, blurring of the observed image of the cells to which vibration has been applied may occur when the cells are observed. [Means for solving the problem]

[0005] a cell detachment device that detaches cells provided on a culture surface of a culture vessel by applying vibration to the culture vessel; an imaging device having an illumination unit that irradiates the cells with light and an imaging unit that images the cells; 1. An observation system comprising: the illumination unit periodically repeats light emission and extinction, The observation system is characterized in that the duration of the light emission is shorter than the period of the vibration. [Effects of the Invention]

[0006] According to the present disclosure, it is possible to provide a cell observation system that applies vibration to detach cells, and that can observe the cells while reducing blurring of the observed image of the cells to which vibration has been applied. [Brief explanation of the drawings]

[0007] [Figure 1] FIG. 1 is a diagram illustrating an observation system according to the present disclosure. [Figure 2] FIG. 1 is a diagram illustrating an observation system according to the present disclosure. [Figure 3] FIG. 1 is a diagram showing an example of an observation system that employs a microscope as an imaging device. [Figure 4] FIG. 1 is a diagram illustrating a first embodiment. [Figure 5] FIG. 1 is a diagram illustrating a first embodiment. [Figure 6] FIG. 1 is a diagram illustrating a first embodiment. [Figure 7] FIG. 10 is a diagram illustrating a second embodiment. [Figure 8] FIG. 10 is a diagram illustrating a second embodiment. [Figure 9] FIG. 10 is a diagram illustrating a third embodiment. [Figure 10] FIG. 10 is a diagram illustrating a fourth embodiment. [Figure 11] FIG. 10 is a diagram illustrating a fourth embodiment. [Figure 12] FIG. 10 is a diagram illustrating a fifth embodiment. [Figure 13] FIG. 10 is a diagram illustrating a fifth embodiment. [Figure 14] FIG. 10 is a diagram illustrating a sixth embodiment. [Figure 15] FIG. 10 is a diagram illustrating a sixth embodiment. [Figure 16] FIG. 13 is a diagram illustrating a seventh embodiment. [Figure 17] FIG. 13 is a diagram illustrating an eighth embodiment. [Figure 18] FIG. 13 is a diagram illustrating a ninth embodiment. [Figure 19] FIG. 19 is a diagram illustrating a tenth embodiment. [Figure 20] FIG. 19 is a diagram illustrating an eleventh embodiment. [Figure 21] FIG. 22 is a diagram illustrating a twelfth embodiment. [Figure 22] FIG. 22 is a diagram illustrating a twelfth embodiment. [Figure 23] FIG. 22 is a diagram illustrating a twelfth embodiment. [Figure 24] FIG. 22 is a diagram illustrating a thirteenth embodiment. [Figure 25] FIG. 22 is a diagram illustrating a fourteenth embodiment. [Figure 26] FIG. 20 is a diagram illustrating a fifteenth embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0008] The present inventors have investigated a method for observing cells to which vibrations have been applied. As a result, they have found that blurring of the observed image is likely to occur when the period of the applied vibrations is shorter than the shutter speed of the imaging device. As a result of further investigation, the present inventors have found that blurring of the observed image of the cells can be reduced by irradiating the cells with light by periodically repeating light emission and extinction using an illumination unit, and by shortening the continuous light emission time to be shorter than the period of vibration.

[0009] First, an example of an observation system according to the present disclosure will be described with reference to FIG. The observation system 1000 includes a cell detachment device 1 and an imaging device 124 . The cell detachment device 1 detaches cells 3 provided on the culture surface 100c of the culture vessel 100 by applying vibration to the culture vessel 100. The cell detachment device may or may not include the culture vessel 100.

[0010] The image capturing device 124 has an illumination unit 124b that irradiates the cells 3 with light and an image capturing unit 124a that captures images of the cells 3.

[0011] In the example of Figure 1, a vibration tank 107 is provided adjacent to the cell detachment device 1, and the vibration tank 107 is filled with a vibration transmission liquid (not shown) such as water, glycol, or glycerin. A vibration plate 102 to which a piezoelectric element 117 that generates ultrasonic vibrations is bonded is installed on the bottom of the vibration tank 107 and sealed with a rubber packing 120 or the like. A storage tank 108 is arranged next to the vibration tank 107, and is formed in a groove shape so that the vibration transmission liquid can move freely through a liquid path 111.

[0012] A sinking weight 103 is provided to enable the vibration-transmitting liquid to move from the vibration tank 107 through the liquid path 111 to the reservoir tank 108 and vice versa. The weight 103 moves by rotating around a rotation center 114. In the example of Figure 1, the cell detachment device 1 also has an O-ring 121 to prevent water from entering.

[0013] As shown in FIG. 1 , the cell detachment device 1 can be provided with an observation window 116. The cells 3 on the culture vessel 100 can be illuminated from below the culture vessel 100 by an imaging device 124 using an illumination unit 124b and photographed by an imaging unit 124a through the observation window 116. The observation window 116 only needs to have a gap through which the cells 3 in the culture vessel 100 can be photographed. A transparent plate such as a glass plate or a resin plate may be installed, or nothing may be installed. However, depending on the imaging method and arrangement of the imaging device 124, it may not be necessary to photograph the cells 3 from below the culture vessel 100. In that case, the observation window 116 may not be necessary, or may be installed somewhere other than below the culture vessel 100. A typical example of the culture vessel 100 is a dish.

[0014] The observation system 1000 in Fig. 1 shows an example in which the imaging unit 124a and illumination unit 124b of the image capturing device 124 are both located below the culture vessel 100. In this example, the diaphragm 102 is a transparent glass plate to which a piezoelectric element 117 is adhesively bonded, and the imaging unit 124a and illumination unit 124b capture images of cells 3 in the culture vessel 100, which consists of a culture vessel lid 100a and a culture vessel receptacle 100b, through the observation window 116 and the diaphragm 102. In particular, the range illuminated by the illumination unit 124b is limited to an illumination range 1161 within the transparent glass plate that constitutes the observation window 116 and the diaphragm 102, thereby reducing noise and making it easier to obtain clearer images. 122 denotes a partition plate (water level adjustment plate), and 125 denotes a communication amplification processing circuit.

[0015] 2 shows another example of an observation system 1000. Reference numeral 101 denotes a housing, 102 denotes a vibrating plate, 103 denotes a sinking weight, 104 denotes cooling fins, 105 denotes a water pipe, 106 denotes a bottom plate, 107 denotes a vibration tank (the space above the vibrating plate 102), 108 denotes a storage tank, 109 denotes a connector (for power supply and communication), 111 denotes a liquid path, 112 denotes a spirit level, and 120 denotes a rubber packing. The spirit level 112 is provided to check whether the installation location is even or to adjust the inclination. In FIG. 2, the illumination unit 124b is an LED 124b1 (also referred to as an LED light) and is included in the imaging device 124.

[0016] The image capturing device 124 is not necessarily dedicated to the observation system 1000, and may be a microscope such as an inverted microscope. FIG. 3 shows an example in which a microscope is used as the image capturing device 124 of the observation system 1000. Reference numeral 217 denotes a microscope table, and reference numeral 215 denotes a protruding portion that serves as a guide when placing the cell detachment device 1 on the table 217. The cell detachment device 1 has a transparent diaphragm 102 disposed below the culture vessel 100. In the example of FIG. 3, the lens 124a2 is an objective lens attached to a revolver, which moves along a rotational trajectory indicated by an arrow in FIG. 3 and can be fixed at a desired position and focused. Cells 3 (not shown in FIG. 3) in the culture vessel 100 set in the cell detachment device 1 placed on the table 217 can be observed through the lens 124a2 by the microscope (not shown in its entirety in FIG. 3), which is the image capturing device 124.

[0017] First embodiment A first embodiment of the present disclosure will be described with reference to FIGS. 1, 4, 5, and 6. FIG. The observation system 1000 of the present disclosure includes a cell detachment device 1 and an imaging device 124. The cell detachment device 1 detaches cells 3 provided on the culture surface 100c of the culture container 100 by applying vibration to the culture container 100. The imaging device 124 has an illumination unit 124b that irradiates light onto the cells 3 and an imaging unit 124a that images the cells 3. The illumination unit 124b periodically repeats light emission and extinction.

[0018] In the first embodiment, the light emission of the illumination unit 124b is linked to the voltage for light emission. When the illumination unit 124b is an LED 124b1 that emits light only when a voltage is applied in the forward direction and the power supply is turned on with an alternating voltage, the LED 124b1 repeatedly emits and extinguishes light in a short cycle. The inventors have noticed that the cycle of this emission and extinguishing (sometimes called the emission cycle) can be made to be approximately the same as the cycle of the vibration (sometimes simply called vibration or detachment vibration) applied to the culture vessel by the cell detachment device 1 to detach the cells 3. In Figure 4, the vertical axis represents the detachment vibration (top) or the emission and extinguishing pattern of the illumination unit 124b (bottom), and the horizontal axis represents time. The detachment vibration is a continuous sine wave. The illumination unit 124b periodically emits and extinguishes light.

[0019] As shown in FIG. 4, in this embodiment, the period of the peeling vibration and the period of the light emission coincide with each other. If the period of the peeling vibration is T1 and the light emission period of the illumination unit 124b is T2, then It is preferable to satisfy at least one of T1=T2×n1 (n1 is an integer) (formula 1) or T1×n2=T2 (n2 is an integer) (formula 2).

[0020] In this embodiment, both formula 1 and formula 2 are satisfied, and n1 = n2 = 1. Furthermore, the continuous light emission time TL is shorter than the period T1 of the peeling vibration. Since the above is satisfied, the illumination unit 124b emits light only when the amplitude of the detachment vibration is within a specific range, which reduces blurring of the observed image of the cell 3 and enables the cell 3 to be photographed clearly.

[0021] 5, an LED 124b1 is used as the lighting unit 124b, and by controlling the LED 124b1 and the piezoelectric element 117 (not shown in FIG. 5) with an alternating voltage synchronization control circuit 123, it is possible to synchronize the light emission cycle of the lighting unit 124b with the peeling vibration cycle of the diaphragm 102. By triggering the rising edge of the drive voltage of the voltage generation unit with a comparator or the like, a voltage (LED voltage) for emitting light from the LED 124b1 can be generated and applied to the LED 124b1.

[0022] 6 conceptually illustrates how cells 3 are photographed while being detached in the observation system of the first embodiment. A culture vessel 100 is placed on a diaphragm 102, and cells 3 are adhered to the culture surface located at the bottom of the culture vessel 100. In this example, an illumination unit 124b is disposed diagonally upward, and an imaging unit 124a is disposed on the underside of the diaphragm 102. In this example, the imaging unit 124a includes a lens 124a2 and an imaging device 124a1. The arrangement of the lens 124a2, imaging device 124a1, and illumination unit 124b can be selectively combined as needed depending on the location and manner of use.

[0023] The concept when the amplitude of the detachment vibration is close to the minimum value in Figure 4 (when it is (a) in Figure 4) is shown in (a) of Figure 6, and the concept when the amplitude of the detachment vibration is close to the maximum value (when it is (b) in Figure 4) is shown in (b) of Figure 6. In (a), the amplitude of the detachment vibration is close to the minimum value and is rising. At this time, the cell 3 is pressed against the culture vessel 100, and a force is applied in the direction shown by the arrow in the figure. The illumination unit 124b is extinguished.

[0024] (b) shows the state when the amplitude of the detachment vibration is close to its maximum value and the position of the culture vessel changes from rising to falling (bottom stop point). At this time, a force acts to separate the cell 3 from the culture vessel 100, causing the cell 3 to float up and the end of the cell 3 to be pulled and stretched. At this time, the illumination unit 124b is emitting light.

[0025] While the peeling vibration is applied, (a) and (b) are repeated. During this time, an image is captured by the image capturer 124a1 only when the illumination unit 124b emits light, that is, the state of the cell 3 in (a) floating up is selectively captured. As a result, blurring of the observed image of the cell 3 is reduced, and the cell 3 can be captured clearly.

[0026] 6(c) shows the state where the peeling is completed. When the completion of the peeling is detected, the illumination unit 124b is turned off.

[0027] The above has described how cells 3 arranged on the culture surface of the culture vessel 100 are detached by applying vibration to the culture vessel 100, and how during this process the illumination unit 124b repeatedly emits and extinguishes light in accordance with the cycle of the detachment vibration of the cells 3, and the cells are selectively photographed by the image capture device 124a1.

[0028] Second embodiment The second embodiment will be described with reference to FIGS. In the first embodiment, the light emission period is the same as the period of the peeling vibration, and the peeling direction is vertical. In the second embodiment, as shown in FIG. 7, the light emission period is half the period of the peeling vibration, and the peeling direction is horizontal. That is, if the period of the peeling vibration is T1 and the light emission period is T2, then T1 = T2 × 2. The continuous light emission time TL is shorter than the period T1 of the peeling vibration.

[0029] In Fig. 8, (a) shows the state before and after the movement of the position of the culture vessel 100 changes from left to right due to the detachment vibration (when it is (a) in Fig. 7). At this time, a rightward force (indicated by the arrow) acts to separate the cell 3 from the culture vessel 100, and in the figure, the cell 3 is being pulled. At this time, the illumination unit 124b emits light.

[0030] The diagram (b) shows the state before and after the detachment vibration causes the position of the culture vessel 100 to shift from right to left (when it is at (b) in FIG. 7). At this time, a leftward force (indicated by the arrow) acts to separate the cell 3 from the culture vessel 100, pulling the cell 3. At this time, the illumination unit 124b emits light. In this way, the direction of the detachment vibration may be a direction other than vertical.

[0031] As described above, the state of the cell 3 in the pulled state is selectively photographed, and as a result, blurring of the observed image of the cell 3 is reduced, and the cell 3 can be photographed clearly.

[0032] Third embodiment FIG. 9 shows an example in which T1×n2=T2 (n2 is an integer) (Equation 2) is satisfied, where T1 is the period of the detachment vibration and T2 is the light emission period. In this example, n2=2. The continuous light emission time TL is shorter than the period T1 of the detachment vibration. In this case, too, the illumination unit 124b emits light when the direction of movement of the culture vessel 100 changes, so that the state in which the cell 3 is lifted is selectively captured. As a result, blurring of the observed image of the cell 3 is reduced, and the cell 3 can be captured clearly.

[0033] Fourth embodiment The fourth embodiment will be described with reference to FIGS. In this embodiment, as shown in FIG. 10, the continuous light emission time TL of the illumination unit 124b is significantly shorter than the period T1 of the peeling vibration.

[0034] FIG. 11 shows, in the order of (a) to (f), how the cell 3 gradually peels off as the time during which the peeling vibration is applied passes.

[0035] In (a), (c), and (e), the position of the culture vessel is rising (when it is (a) in Figure 4), and (b), (d), and (f) show the state before and after the position of the culture vessel changes from rising to falling (when it is (b) in Figure 4).

[0036] An image is captured by the image capturer 124a1 only when the illumination unit 124b emits light, i.e., the state of the cell 3 floating in (b), (d), and (f) is selectively captured, reducing blurring of the observed image of the cell 3 and enabling the cell 3 to be captured clearly.

[0037] Fifth embodiment The fifth embodiment will be described with reference to FIGS. Generally, with high-speed cameras and high-speed frame cameras, there is a trade-off between image quality and the imaging speed per frame. For example, capturing a cell 3 approximately 10 μm or less in size in a culture vessel 100 requires only a few pixels, even with a field of view diameter of 5 mm and several megapixels. Therefore, to obtain a clear image, the imaging speed must be reduced.

[0038] The present disclosure is useful in such situations. As disclosed in Patent Document 1 and the like, there are various ways to capture cell images.

[0039] It is effective to suppress blurring by periodically repeating light emission and extinction of the illumination unit 124 while shortening the continuous light emission time TL for each period. In this embodiment, as shown in Fig. 12, the continuous irradiation time TL is set to 50000 ns or less.

[0040] As shown in Figure 13, the illumination unit 124 periodically emits and extinguishes light, but if the continuous irradiation time TL is long, as in the illumination unit 124 shown below, the images of cells 3 will overlap with each other, as shown in the lower right of the figure. When the illumination unit 124 periodically turns on and off, and the continuous irradiation time TL is set to 50,000 ns or less, as in the illumination unit 124 shown above, the image of cell 3 is clear with little overlap between cells, as shown in the upper right of the figure.

[0041] Sixth embodiment The sixth embodiment will be described with reference to FIGS. In this embodiment, the phase of the light emission cycle can be changed. In this embodiment, the light emission cycle of the illumination unit 124 is out of phase with the cycle of the peeling vibration. In FIG. 14(a), the phases of the peeling vibration and the light emission cycle of the illumination unit 124 match, and the peaks overlap. On the other hand, in (b), the phases of the peeling vibration and the light emission cycle of the illumination unit 124 are out of phase, and the peaks are out of phase. In (b), light is emitted at the bottom of the vibration, that is, at the timing when the culture vessel 100 changes from descending to ascending. The way in which the phase is changed can be determined according to the desired image.

[0042] The adjustment of the peeling vibration and the vibration periods of the illumination unit 124b and diaphragm 102 was described above with reference to FIG. 5. A means for changing the phase can be provided by providing a delay circuit in the synchronization control circuit. As an example, the phase can be shifted arbitrarily by varying the comparator voltage. The phase can also be selected by providing a switch for changing the voltage value on the GUI monitor 300 as shown in FIG. 15, or by providing a voltage dividing resistor or the like to a predetermined voltage dividing value. Reference numeral 123 denotes the synchronization control circuit, 1501 denotes a phase button, and 1502 denotes an animation button.

[0043] Seventh embodiment The first embodiment has shown an example of a means for synchronizing the cycles of the peeling vibration and the illumination unit 124b. The seventh embodiment provides a means for adjusting the cycles of the peeling vibration and the illumination unit 124b other than the driving alternating voltage.

[0044] There is a system that can sense and monitor the vibration of the diaphragm 102. A control device 200 will be described with reference to FIG. 16. The control device 200 according to this embodiment includes a sensor voltage detection unit 201, a vibration control unit 202, a determination unit 203, a notification control unit 204, and a storage unit 205. The sensor voltage detection unit 201 is connected to the sensor electrode 5 and the ground electrode 6. The sensor voltage detection unit 201 can detect a voltage generated in the piezoelectric element 117 when vibrations of the vibration-receiving system, including the diaphragm 102, the culture vessel 100, the vibration transmitter, and the like, which receive vibrations generated by the piezoelectric element 117, are applied to the piezoelectric element 117.

[0045] At this time, the detected voltage is compared with the comparator to obtain the LED voltage as described in embodiment 1. Also, an LED voltage according to the photographing phase condition designated by the user can be applied to the LED 124b1.

[0046] In addition, the vibration control unit 202 can also be connected to a GUI monitor 300 equipped with, for example, a touch panel, and the user can input shooting conditions and give instructions to start and stop vibration and light emission via the display screen of this GUI monitor 300.

[0047] The determination unit 203 performs processing such as AD conversion on the voltage generated by the piezoelectric element 117 detected by the sensor voltage detection unit 201, and can determine whether the vibration occurring in the vibration system described above at the time of detection is appropriate based on the obtained voltage value. The notification control unit 204 is connected to the GUI monitor 300 and can display the determination result of the determination unit 203 on the GUI monitor 300 in a predetermined format. The storage unit 205 can store, for example, the determination criteria of the determination unit 203 based on the relationship between the input current to the drive electrode 7 and the voltage value output by the sensor electrode 5 corresponding to the input current, as well as a program for operating the notification control unit 204 in accordance with the determination result of the determination unit 203. For example, the LED 124b1 may be illuminated based on this determination value to record the occurrence of an abnormal amplitude state. The imaging conditions of the illumination unit 124b and the imaging unit 124a may be linked to the driving conditions for each cell type.

[0048] In the illustrated example, the control device 200 and the GUI monitor 300 are shown as separate components, but they may be integrated into a personal computer, or a tablet personal computer or other portable terminal may be used instead.

[0049] As described above, the illumination unit 124b can change the phase depending on the vibration state of the cell detachment device 1 or the type of the cell 3.

[0050] Eighth embodiment As an eighth embodiment, an example of adjusting the continuous light emission time TL of the LED 124b1 will be described with reference to Fig. 17. In this embodiment, the LED 124b1 is driven by an alternating voltage.

[0051] The power supply is an AC voltage as shown in (b) of Fig. 17, and a threshold value (shown by a dashed line in the figure) is set for comparing the AC voltage with a comparator. Then, LED 124b1 emits light only when the AC voltage exceeds the threshold value, and the LED 124b1 repeatedly emits and extinguishes light as shown in (a). In this way, by alternately switching and comparing the input voltage of the illumination unit 124b, it is possible to adjust the continuous light emission time of the illumination unit 124b.

[0052] Ninth embodiment As a ninth embodiment, an example in which the imaging unit 124a also uses a shutter, and the shutter repeats a light emission cycle to adjust the light emission cycle, will be described with reference to FIG.

[0053] In Figure 18, the light emission and extinction patterns of the illumination unit 124b are shown at the top, the peeling vibration is shown in the middle, and the electronic shutter or frame data capture cycle is shown at the bottom. In the example of Figure 18, two cycles of peeling vibration occur when the electronic shutter is opened once. In this embodiment, the light emission cycle is four times the peeling vibration cycle so that one light emission occurs in accordance with the shutter cycle. In this way, one light emission occurs per shutter opening. This allows the image capture device 124a1 to capture frames continuously without any extraneous light entering.

[0054] In this case, if the cycle of light emission and extinction of the illumination unit 124b is T2 and the cycle of opening and closing the shutter is T3, it is preferable that T2 is an integer multiple of T3, or that T3 is an integer multiple of T2. In other words, T2 and T3 satisfy the relationship of at least one of Equation 3 and Equation 4. T2 = T3 × n3 (n3 is an integer) (Equation 3) T2×n4=T3 (n4 is an integer) (Equation 4)

[0055] Tenth embodiment As a tenth embodiment, with reference to FIG. 19, an example in which an illumination unit 124b includes an LED 124b1 as a light source will be described.

[0056] The LED 124b1 periodically turns on and off in response to the LED voltage. In Figure 19, the vertical axis represents the LED voltage, and the horizontal axis represents the light emission response time of the LED 124b1. The LED voltage has a short cycle, allowing for a response at about 5 MHz, which can be set to 100 ns or less. As a result, the light emission cycle of the LED 124b1 can be set to 50 to 100 ns.

[0057] Eleventh embodiment As an eleventh embodiment, with reference to FIG. 20, an example in which a general camera is used as the imaging unit 124a will be described.

[0058] In Figure 20, the shutter open time is shown at the top, the peeling vibration is shown in the middle, and the light emission and extinction pattern of the illumination unit 124b is shown at the bottom. As shown in Figure 20, the shutter open time of a commercially available camera is longer than the peeling vibration. Therefore, when using a general camera as the imaging unit 124a, it is recommended that multiple light emission cycles be included in one shutter open time.

[0059] In this example, if the cycle of light emission and extinction of the illumination unit 124b is T2 and the cycle of opening and closing the shutter is T3, T2 and T3 are T3=T2×6, that is, this is an example where N4=6 in equation 4.

[0060] This allows a large amount of light to be obtained in overlapping fashion, with little blurring, making it easier to observe easily and accurately using a general camera.

[0061] Twelfth embodiment As a twelfth embodiment, an example of an observation system for capturing an image of an animation portion of a cell 3 will be described with reference to FIGS.

[0062] FIG. 21 shows the process of peeling a cell 3 using peeling vibration, in which the illumination unit 124b periodically turns on and off while photographing the cell 3, with time passing in the order of (a), (b), and (c).

[0063] The appearance of cell 3 is the same as that explained for Figure 6. In Figure 21, the right side shows images of cell 3 taken at (a), (b), and (c) respectively. In the center of Figure 21, it is explained that imaging is repeated for each illumination cycle, and that when the illumination cycle is repeated N times, N images are obtained. By accumulating these images, the peeling of cell 3 can be animated.

[0064] The imaging system of this embodiment can include a GUI monitor 300, as shown in Fig. 22. An enlarged view of the GUI monitor 300 in Fig. 22 is shown in Fig. 23. The imaging system has an imaging position selection button 2302 for selecting the position of the culture vessel during imaging. Furthermore, the imaging system of this embodiment can have an animation button 2301 for selecting an animation.

[0065] Thirteenth embodiment As a thirteenth embodiment, an observation system having a determination unit for a cell 3 is shown with reference to FIG. In the observation system of the present disclosure, the illumination unit 124b periodically turns on and off, so that even if there is detachment vibration, the cells 3 can be photographed when the culture vessel 100 is in the same position, making it easy to compare images of the cells 3. Fig. 24 shows an example of a GUI monitor 300 that can be included in the observation system of this embodiment.

[0066] When it is desired to reproduce the irradiation position at the time when the characteristics of each cell type were captured, this can be memorized using the memory button 2401 in Fig. 24. In Fig. 24, the rising point of the imaging position selection button 2302 is flashing, showing an example in which the rising point has been memorized as the imaging position. Furthermore, the drive frequency, amplitude voltage, LED voltage, etc. can be recorded all at once, and these can be recalled using the observation condition button 2402 or recall button 2403, making it possible to observe under the previously recorded conditions.

[0067] In this way, by appropriately setting the timing of light emission and the cell observation conditions, it is possible to determine whether or not the detachment of the cell 3 is complete. Furthermore, information on the state of the cell 3, such as the state of damage to the cell 3, can be obtained.

[0068] Fourteenth embodiment As a fourteenth embodiment, referring to FIG. 25, an observation system is shown in which the imaging unit 124a can be moved to a retreat position where it does not interfere with the installation, removal, etc. of the incubation container 100.

[0069] FIG. 25 shows the process of peeling cell 3 by peeling vibration, with the illumination unit 124b periodically repeating light emission and extinction while photographing cell 3, with time passing in the order of (a), (b), and (c). The state of cell 3 during the peeling process is the same as that explained in FIG. 6. In FIG. 25, (a) and (b) show the cell 3 being peeled, and

[0070] (a) shows the state during ascent, and (b) shows the state before and after the bottom stop point. During this period, the image capture device 124 is above the culture vessel 100, i.e., above the cell detachment device 1. On the other hand, in (c) showing the state after the cell 3 has been detached, the image capture device 124 has moved to a retracted position that is not above the culture vessel 100, as indicated by the diagonal arrow. The movement is preferably a relative movement with respect to the culture vessel 100. The ability of the image capture unit 124a to move to the retracted position makes it easy to remove the culture vessel 100. Note that the image capture device 124 can be moved to the retracted position whenever desired, not only after detachment but also before or during detachment.

[0071] Fifteenth embodiment As a fifteenth embodiment, a method for producing cells is provided, as shown in FIG. The cell production method of this embodiment includes a culture step S2601 and a detachment step S2602. In the culture step S2601, cells are cultured on the culture surface of a culture vessel. In the detachment step S2602, the cells are detached from the culture surface by applying vibration to the cells. Furthermore, in the detachment step, the cells are irradiated with light so that they are periodically irradiated, and if the period of vibration is T5 and the period of cell irradiation is T6, T5 and T6 satisfy the relationship of at least one of Equation 5 and Equation 6. T5 = T6 × n5 (n5 is an integer) (Equation 5) T6×n6=T5 (n6 is an integer) (Equation 6)

[0072] The disclosure of this embodiment includes the following configurations and methods. (Configuration 1) a cell detachment device that detaches cells provided on a culture surface of a culture vessel by applying vibration to the culture vessel; an imaging device having an illumination unit that irradiates the cells with light and an imaging unit that images the cells; 1. An observation system comprising: the illumination unit periodically repeats light emission and extinction, An observation system characterized in that the continuous emission time of the light is shorter than the period of the vibration. (Configuration 2) The observation system described in configuration 1 is characterized in that, when the period of the vibration is T1 and the period of light emission and extinction of the illumination unit is T2, T1 and T2 satisfy the relationship of at least one of formulas 1 and 2. T1 = T2 × n1 (n1 is an integer) (Equation 1) T1×n2=T2 (n2 is an integer) (Equation 2) (Configuration 3) 3. The observation system according to any one of configurations 1 and 2, wherein the continuous emission time of the light is 50,000 ns or less. (Configuration 4) 4. The observation system according to any one of configurations 1 to 3, wherein the phase of the optical cycle, which is the period during which the illumination unit periodically repeats light emission and extinction, is different from the phase of the vibration. (Configuration 5) The observation system described in any one of configurations 1 to 4, characterized in that the illumination unit is capable of changing the phase of the light emission cycle depending on the vibration state of the cell detachment device or the type of cell. (Configuration 6) 6. The observation system according to any one of configurations 1 to 5, wherein the illumination unit is driven by an alternating voltage. (Configuration 7) 7. The observation system according to any one of configurations 1 to 6, wherein the illumination unit includes an LED. (Configuration 8) 8. The observation system according to any one of configurations 1 to 7, wherein the imaging unit has a shutter that periodically repeats opening and closing. (Configuration 9) The observation system described in configuration 8 is characterized in that, when the period of light emission and extinction of the illumination unit is T2 and the period of opening and closing of the shutter is T3, T2 and T3 satisfy the relationship of at least one of Equation 3 and Equation 4. T2 = T3 × n3 (n3 is an integer) (Equation 3) T2×n4=T3 (n4 is an integer) (Equation 4) (Method 1) A culturing step of culturing cells on the culture surface of a culture vessel; and A cell manufacturing method including a detachment step of detaching the cells from the culture surface by applying vibration to the cells, In the detachment step, light is irradiated so that the cells are periodically irradiated; A method for producing cells, characterized in that, when the period of the vibration is T5 and the period during which the cells are irradiated is T6, T5 and T6 satisfy the relationship of at least one of formulas 5 and 6. T5 = T6 × n5 (n5 is an integer) (Equation 5) T6×n6=T5 (n6 is an integer) (Equation 6). [Explanation of symbols]

[0073] 1. Cell detachment device 3 cells 100 culture vessels 100c culture surface 124 Imaging Device 124a Imaging unit 124b Lighting section 124b1 LED TL Continuous light emission time

Claims

1. a cell detachment device that detaches cells provided on a culture surface of a culture vessel by applying vibration to the culture vessel; an imaging device having an illumination unit that irradiates the cells with light and an imaging unit that images the cells; 1. An observation system comprising: the illumination unit periodically repeats light emission and extinction, An observation system characterized in that the continuous emission time of the light is shorter than the period of the vibration.

2. 2. The observation system according to claim 1, wherein, when the period of the vibration is T1 and the period of light emission and extinction of the illumination unit is T2, T1 and T2 satisfy the relationship of at least one of Equation 1 and Equation 2. T1 = T2 × n1 (n1 is an integer) (Equation 1) T1 × n2 = T2 (n2 is an integer) (Equation 2)

3. 2. The observation system according to claim 1, wherein the light emission duration is 50,000 ns or less.

4. 2. The observation system according to claim 1, wherein the phase of a light cycle, which is a period during which the illumination unit periodically repeats light emission and extinction, is different from the phase of the vibration.

5. 2. The observation system according to claim 1, wherein the illumination unit is capable of changing the phase of the light emission cycle depending on the state of the vibration of the cell dissociation device or the type of the cell.

6. 2. The observation system according to claim 1, wherein the illumination unit is driven by an alternating voltage.

7. 2. The observation system of claim 1, wherein the illumination unit includes an LED.

8. 2. The observation system according to claim 1, wherein the imaging unit has a shutter that periodically repeats opening and closing.

9. The observation system according to claim 8, wherein when the period of light emission and extinction of the illumination unit is T2 and the period of opening and closing of the shutter is T3, T2 and T3 satisfy the relationship of at least one of Equation 3 and Equation 4. T2 = T3 × n3 (n3 is an integer) (Equation 3) T2 × n4 = T3 (n4 is an integer) (Equation 4)

10. A culturing step of culturing cells on the culture surface of a culture vessel; and A cell manufacturing method including a detachment step of detaching the cells from the culture surface by applying vibration to the cells, In the detachment step, light is irradiated so that the cells are periodically irradiated; A method for producing cells, characterized in that, when the period of the vibration is T5 and the period during which the cells are irradiated is T6, T5 and T6 satisfy the relationship of at least one of formulas 5 and 6. T5 = T6 × n5 (n5 is an integer) (Equation 5) T6 × n6 = T5 (n6 is an integer) (Equation 6)

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