In vitro experimental system and temperature regulation method thereof

Abstract

The present invention provides an in vitro experimental system, comprising: an incubator, the incubator comprising a temperature regulator for regulating the internal temperature of the incubator; a culture device, arranged inside the incubator, comprising: at least one culture dish for containing culture fluid for culturing biological tissues, and at least one group of electrode plates for applying an electric field to the culture fluid contained in the corresponding culture dish; and a temperature detector for detecting the temperature of the culture fluid contained in at least one culture dish, wherein the temperature regulator is used to adjust the internal temperature of the incubator so that the temperature of the culture fluid falls within the temperature threshold range when the temperature detector detects that the temperature of the culture fluid exceeds the temperature threshold range. The in vitro experimental system of the present invention adjusts the internal temperature of the incubator by adjusting the temperature regulator of the incubator so that the temperature of the culture fluid falls within the temperature threshold range, without adjusting the alternating voltage, thereby avoiding the influence of the fluctuation of the field strength on the experiment.

Description

Technical Field

[0001] The present invention relates to the field of in vitro experimental equipment, and in particular to an in vitro experimental system and a temperature regulating method thereof. Background Art

[0002] Tumor Treatment Fields (TTFields) is a treatment method that inhibits tumor cell proliferation by interfering with tumor cell mitosis. This low-intensity, medium-high frequency alternating electric field is used to intervene in the treatment of a variety of tumor diseases and is a clinically effective cancer treatment method. Many basic studies at home and abroad have confirmed the biological effects of TTFields. The results of cell experiment studies have shown that TTFields can regulate the expression of biochemical molecules in tumor cells, interfere with the division cycle of tumor cells, inhibit the proliferation of tumor cells and promote tumor cell apoptosis. The results of animal experiment studies have shown that TTFields can effectively inhibit tumor angiogenesis and inhibit the growth of tumors in tumor-bearing mice. The current clinical research results at home and abroad have also confirmed the positive effect of TTFields on tumor treatment.

[0003] In in vitro studies, TTFields will generate a large amount of heat energy when acting on the cell culture fluid in the cell culture device, causing the temperature of the culture fluid to continue to rise. However, the in vitro culture of biological tissues has high requirements for the environmental temperature, and the cell culture fluid needs to be maintained at around 37°C, otherwise the biological tissue is prone to apoptosis due to inappropriate environmental temperature, affecting the reference of TTFields in vitro experiments. The temperature of the culture fluid disclosed in the existing invention disclosure patent No. 112680349 is monitored by a thermistor set on the electrode plate. The temperature signal monitored by the thermistor is transmitted to the external control unit. The external control unit adjusts the output intensity of the alternating current signal according to the temperature signal to control the heat generated by the electric field, so that the temperature of the cell culture fluid is maintained at around 37°C. The above-mentioned method of maintaining the temperature of the cell culture fluid is achieved by adjusting the intensity of the electrical signal output, causing fluctuations in the strength of the alternating electric field. This fluctuating field strength is not conducive to the repeatability of the in vitro research experiment itself and the quantitative statistics of the experimental results. At the same time, the thermistor on the electrode sheet is immersed in the culture medium for a long time, which can easily cause the cell culture medium to penetrate into the sealing glue covering the outer periphery of the thermistor and affect the thermistor, resulting in errors in the temperature monitored by the thermistor and further affecting the temperature of the culture medium.

[0004] Therefore, it is indeed necessary to provide an improved in vitro experimental system and a temperature regulation method thereof to avoid the deficiencies of the above-mentioned in vitro experimental system and temperature regulation method. Summary of the invention

[0005] The invention provides an in vitro experimental system with stable field intensity and accurate temperature monitoring and a temperature regulating method thereof.

[0006] The in vitro experimental system of the present invention is implemented through the following technical scheme: an in vitro experimental system, comprising: an incubator, the incubator comprising a temperature regulator, the temperature regulator being configured to adjust the internal temperature of the incubator; a culture device, arranged inside the incubator, comprising: at least one culture dish for containing culture fluid for culturing biological tissue; and at least one group of electrode plates, each group of electrode plates being at least partially arranged in a corresponding culture dish of the at least one culture dish, for applying an electric field to the culture fluid contained in the culture dish; and a thermometer, for detecting the temperature of the culture fluid contained in the at least one culture dish, wherein the temperature regulator adjusts the internal temperature of the incubator so that the culture fluid temperature falls within the temperature threshold range when the thermometer detects that the temperature of the culture fluid exceeds a temperature threshold range.

[0007] Furthermore, the temperature measuring device includes: at least one temperature measuring line, each temperature measuring line has a thermocouple probe at the end, and the thermocouple probe extends into the culture liquid in the corresponding culture dish; and a test instrument for converting the at least one detection signal into a corresponding temperature value.

[0008] Furthermore, the culture device also includes: a circuit board, which is arranged above the at least one culture dish and is used to electrically connect at least one group of electrode plates.

[0009] Furthermore, the circuit board is provided with a plurality of sockets for plugging in the at least one group of electrode plates, and at least one pinhole for passing the temperature measuring line is also provided in the area surrounded by the plurality of sockets. The at least one culture dish is provided with a corresponding opening at its respective top end. The at least one group of electrode plates is arranged on the circuit board through the plurality of sockets, and extends into the at least one culture dish through the corresponding opening of the at least one culture dish in a direction away from the circuit board.

[0010] Furthermore, each electrode plate group includes at least two pairs of electrode plates.

[0011] Furthermore, each electrode plate includes a flexible circuit board, an insulating substrate and a ceramic electrode arranged on opposite sides of the flexible circuit board.

[0012] Furthermore, the flexible circuit board and the insulating substrate have the same shape, and are both arranged in a "T" shape.

[0013] Furthermore, the width of the flexible circuit board is smaller at one end close to the circuit board, while the width of the flexible circuit board is larger at one end away from the circuit board.

[0014] Furthermore, the ceramic electrode is arranged at one end of the flexible circuit board away from the circuit board.

[0015] Furthermore, each electrode plate also includes at least one temperature sensor disposed at one end of the flexible circuit board away from the circuit board, and the temperature sensor is disposed on the same side of the flexible circuit board as the ceramic electrode.

[0016] Furthermore, each electrode plate also includes a plurality of conductive terminals arranged at one end of the flexible circuit board close to the circuit board, one end of each conductive terminal is electrically connected to the flexible circuit board, and the other end is electrically connected to the circuit board above the culture dish.

[0017] Furthermore, the other end of each conductive terminal is electrically connected to a corresponding socket on the circuit board.

[0018] Furthermore, the culture device also includes: a mounting plate assembly for mounting and fixing the at least one culture dish and the circuit board.

[0019] Furthermore, the mounting plate assembly includes a bottom plate, a plurality of support columns fixed on the bottom plate, and a top plate detachably fixed to the plurality of support columns.

[0020] Furthermore, the bottom plate is a plate-shaped structure for supporting at least one culture dish.

[0021] Furthermore, a side surface of the bottom plate supporting the culture dish is provided with at least one positioning component for limiting horizontal movement of the culture dish.

[0022] Furthermore, a first fixing component is provided on the top of each supporting column.

[0023] Furthermore, the top plate is provided with a plurality of second fixing components at positions corresponding to the first fixing components of the plurality of support columns in a one-to-one correspondence.

[0024] Furthermore, the top plate is detachably fixedly connected to the first fixing parts of the corresponding support columns through its multiple second fixing parts.

[0025] Furthermore, the top plate is arranged above at least one culture dish to support the circuit board.

[0026] Furthermore, at least one limiting opening is formed on the top plate, for accommodating the top end of at least one culture dish respectively.

[0027] Furthermore, a flat cable connection terminal is provided at one end of the circuit board, and the flat cable connection terminal is used to connect a flat cable.

[0028] Furthermore, it also includes: an electric field generator, which is used to generate at least one alternating electric field signal and transmit it to at least one group of electrode plates.

[0029] Furthermore, it also includes an adapter, which is electrically connected to the electric field generator and is used to provide the at least one alternating electric field signal to at least one group of electrode plates through the adapter.

[0030] Furthermore, it also includes: a controller, wherein the controller is configured to automatically control the temperature regulator to adjust the internal temperature of the incubator so that the culture solution temperature falls within the temperature threshold range in response to detecting that the culture solution temperature exceeds the temperature threshold range.

[0031] Furthermore, the temperature threshold range is 36°C-38°C.

[0032] The temperature regulation method of the in vitro experimental system of the present invention is achieved through the following scheme: a temperature regulation method of the in vitro experimental system as described above, the method comprising: using the thermometer to periodically detect the temperature of the culture solution contained in the at least one culture dish; and in response to detecting that the temperature of the culture solution exceeds the temperature threshold range, controlling the temperature regulator to adjust the internal temperature of the incubator so that the temperature of the culture solution falls within the temperature threshold range, and the temperature threshold range is 36°C-38°C.

[0033] Furthermore, the temperature threshold range includes an ideal temperature value that the culture solution is expected to maintain, wherein controlling the temperature regulator to adjust the internal temperature of the incubator also includes: obtaining a current operating temperature set for the temperature regulator; and setting a target operating temperature of the temperature regulator based on the difference between the current operating temperature and the culture solution temperature and the ideal temperature value.

[0034] Furthermore, the method further includes: in response to detecting that the culture solution temperature does not exceed the temperature threshold range, not changing the temperature setting of the temperature regulator.

[0035] The in vitro experimental system of the present invention utilizes a thermometer to detect the temperature inside a culture dish. When it is detected that the temperature of the culture solution exceeds a temperature threshold range, the internal temperature of the incubator is adjusted by adjusting the temperature regulator of the incubator so that the temperature of the culture solution falls within the temperature threshold range. There is no need to adjust the alternating voltage, thereby avoiding the influence of fluctuations in the field strength on the experiment, thereby facilitating the repeatability of the experiment itself in in vitro research, quantitative statistics of the experimental results, etc.

[0036] These and other aspects of the disclosure will be apparent from and elucidated with reference to the embodiments described hereinafter. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 is a schematic block diagram of an in vitro experimental system according to the present invention.

[0038] Figure 2 yes Figure 1 A three-dimensional diagram of the culture device and the temperature measuring device in FIG.

[0039] Figure 3 yes Figure 2 Schematic diagram of the AA section of the culture device in;

[0040] Figure 4 yes Figure 2 A three-dimensional exploded view of the culture device in FIG.

[0041] Figure 5 yes Figure 4 A schematic plan view of an electrode plate of a culture device in FIG. 1 ; and

[0042] Figure 6 yes Figure 5 A perspective exploded view of the electrode plate shown;

[0043] Figure 7 yes Figure 4 A three-dimensional diagram of a circuit board of a culture device;

[0044] Figure 8 yes Figure 2 Schematic diagram of the culture device during assembly;

[0045] Fig. 9 is a flow chart of a temperature regulation method for an in vitro experimental system according to a first embodiment of the present invention;

[0046] Fig.10 is a flow chart of a temperature regulation method for an in vitro experimental system according to a second embodiment of the present invention. DETAILED DESCRIPTION

[0047] Exemplary embodiments will be described in detail herein, examples of which are shown in the accompanying drawings. When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present application. Instead, they are merely examples of electrode sheets consistent with some aspects of the present application as detailed in the attached claims.

[0048] Figures 1 to 8The in vitro experimental system 1000 of the present invention is shown, which includes a culture device 100, an incubator 200 for placing the culture device 100, a thermocouple-type temperature detector 300 for monitoring the temperature of the culture solution in the culture device 100, an electric field generator 410 for providing an alternating electric signal, an adapter 420 for transmitting the alternating electric signal from the electric field generator 410 to the culture device, and an air conditioner 500 for controlling the spatial environment temperature (room temperature) of the in vitro experimental system 1. The incubator 200 includes a temperature regulator 210 for regulating the heating temperature of the incubator 200 to regulate the ambient temperature in the incubator 200. The in vitro experimental system 1000 monitors the temperature of the culture solution in the culture device 100 through the thermocouple-type temperature detector 300, and the monitored temperature data is more accurate and reliable. The in vitro experimental system 1000 adjusts the heating temperature of the temperature regulator 210 according to the temperature of the culture fluid monitored by the thermometer 300 so that the temperature of the culture fluid in the culture device 100 placed in the incubator 200 falls within the temperature threshold range of 36°C-38°C, and the air conditioner 500 is used to assist in adjusting the ambient temperature in the incubator 200 so that the heat in the incubator 200 is transferred to the space environment outside the incubator 200, so that the cell culture fluid can be carried out in the TTF in vitro research experiment under the field strength environment of the stable alternating electric field, which is conducive to the repeatability of the in vitro research experiment and the quantitative statistics of the experimental results. In some cases, the air conditioner 500 in the in vitro experimental system 1000 is not necessary, such as when the spatial environment temperature is lower than the temperature in the incubator 200, the heat in the incubator 200 is transferred to the space environment outside the incubator 200. In some cases, the air conditioner 500 in the in vitro experimental system 1000 is necessary. For example, if the spatial environment temperature is higher than the temperature inside the incubator 200, the heat inside the incubator 200 cannot be transferred to the spatial environment outside the incubator 200. It is necessary to use the air conditioner 500 to lower the temperature of the spatial environment outside the incubator 200 until it is lower than the temperature inside the incubator 200, and the heat inside the incubator 200 is then transferred to the spatial environment outside the incubator 200.

[0049] The culture device 100 includes at least one culture dish 110, at least one electrode plate group 120, a mounting plate assembly 130 for fixing at least one culture dish 110, and a circuit board 140 located above at least one culture dish 110 and electrically connected to at least one electrode plate group 120. The culture dish 110 contains a culture solution for culturing biological tissues. The electrode plate group 120 is at least partially disposed in the culture dish 110, and is used to apply an electric field to the culture solution contained in the culture dish 110. In this embodiment, the number of the culture dishes 110 and the electrode plate group 120 can be 3 to form different control groups. Each group of electrode plate groups 120 is disposed in a corresponding culture dish 110. The three culture dishes 110 are arranged in a linear direction at intervals. The size and shape of each culture dish 110 are the same. In this embodiment, each culture dish 110 is a cubic structure, and the culture dish 110 is provided with a corresponding opening 1101 at its top, and the culture solution can be loaded into the culture dish 110 through the opening 1101. In other embodiments, the number of culture dishes 110 may be more than 3 or less than 3, and the culture dishes 110 may also have shapes other than cubes, such as rectangular parallelepiped, cylindrical structure, etc., as well as arrangements other than linear arrangements, which are not listed here one by one.

[0050] Each electrode plate group 120 may include four electrode plates 121, and the four electrode plates 121 form two pairs of electrode plates 121, and each pair of electrode plates 121 includes two electrode plates 121 facing each other and having opposite polarities, so as to apply a uniform electric field to the culture solution. In other embodiments, each electrode plate group 120 may also include other groups of electrode plates 121, and each electrode plate group 120 may also be arranged in other suitable arrangements, which are not listed here one by one.

[0051] Each electrode plate 121 includes a flexible circuit board 1210, an insulating substrate 1211 and a ceramic electrode 1212 disposed on opposite sides of the flexible circuit board 1210. The insulating substrate is used to support the flexible circuit board 1210. The flexible circuit board 1210 and the insulating substrate 1211 have substantially the same shape, and are both substantially arranged in a "T" shape. The width of the end of the flexible circuit board 1210 close to the circuit board 140 is smaller, while the width of the end away from the circuit board 140 is larger, so that the ceramic electrode 1212 is arranged at the end with larger width. There is at least one ceramic electrode 1212 of each electrode plate 121. In this embodiment, the number of ceramic electrodes 1212 of each electrode plate 121 is two. The flexible circuit board 1210 is used to transmit electrical signals. At least one ceramic electrode 1212 is electrically connected to the flexible circuit board 1210 to obtain an electrical signal applying an alternating electric field from the flexible circuit board 1210. In this embodiment, the ceramic electrode 1212 is arranged in a circular sheet shape. The two ceramic electrodes 1212 are welded on the flexible circuit board 1210 by solder (not shown) to achieve electrical connection between the two. A gap (not shown) is generated between the ceramic electrode 1212 and the flexible circuit board 1210 due to welding. The gap (not shown) is also filled with a sealant (not shown) to prevent the ceramic electrode 1212 from being affected by external forces and causing the welding point to break, thereby causing the alternating electric field to be unable to be applied to the cells in the culture medium through the ceramic electrode 1212; at the same time, it can also prevent water vapor in the air from entering the gap (not shown) and corroding the solder (not shown) between the ceramic electrode 1212 and the flexible circuit board 1210, thereby affecting the electrical connection between the ceramic electrode 1212 and the flexible circuit board 1210.

[0052] Each electrode plate 121 is also provided with at least one temperature sensor 1214. The temperature sensor 1214 is provided on the same side of the flexible circuit board 1210 as the ceramic electrode 1212. The temperature sensor 1214 is located at the same end of the ceramic electrode 1212 to facilitate detection of the temperature of the culture solution. The temperature sensor 1214 is coated with a sealant (not shown) around the periphery to protect the temperature sensor 1214. Each electrode plate 121 also includes a group of conductive terminals 1213. The group of conductive terminals 1213 is provided on the end of the flexible circuit board 1210 close to the circuit board 140. One end of the group of conductive terminals 1213 is electrically connected to the flexible circuit board 1210, and the other end is electrically connected to the circuit board 140 above the culture dish 110. The group of conductive terminals 1213 is electrically connected to the ceramic electrode 1212 and the temperature sensor 1214 through the flexible circuit board 1210. In this embodiment, the number of temperature sensors 1214 provided on each electrode plate 121 is 2. Each electrode plate 121 is provided with four conductive terminals 1213 .

[0053] The mounting plate assembly 130 of the culture device 100 is used to mount and fix at least one culture dish 110. The mounting plate assembly 130 includes a bottom plate 131, a plurality of support columns 132 fixed on the bottom plate 131, and a top plate 133 detachably fixed to the plurality of support columns 132.

[0054] The bottom plate 131 is roughly a rectangular plate-shaped structure. The bottom plate 131 is used to support at least one culture dish 110. One side surface of the bottom plate 131 supporting the culture dish 110 is provided with at least one positioning component 1310 for limiting the horizontal movement of the culture dish 110. In this embodiment, three positioning components 1310 are provided. The positioning component 1310 is a groove, and its recessed area is roughly the same shape and size as the bottom surface of the culture dish 110. In other embodiments, the positioning component 1310 is other shapes with the same positioning function, such as a convex ring, a plurality of convex points arranged in an annular manner, etc.

[0055] The bottom ends of the plurality of support columns 132 are fixed on the bottom plate 131, for example, the bottom end of each support column 132 can be fixed on the upper surface of the bottom plate 131 by screws. The plurality of support columns 132 are arranged between two adjacent culture dishes 110 at intervals, and are located at the edges of the opposite sides of the bottom plate 131, so that the plurality of support columns 132 avoid the position of the culture dishes 110, making the overall structure of the culture device 100 more compact. The top of each support column 132 is provided with a first fixing part 1320. Each first fixing part 1320 is arranged in a convex shape on the top of the corresponding support column 132, and each support column 132 is arranged in a substantially "convex" shape. The top plate 133 is provided with a plurality of second fixing parts 1330 corresponding to the first fixing parts 1320 of the plurality of support columns 132. The plurality of second fixing parts 1330 are arranged in a notch shape. The top plate 133 is detachably fixedly connected to the first fixing parts 1320 of the corresponding support column 132 through its multiple second fixing parts 1330. The multiple second fixing parts 1330 of the top plate 133 and the first fixing parts 1320 of the corresponding support column 132 form a mortise and tenon fixation. That is, the first fixing part 1320 is a tenon, and the second fixing part 1330 is a mortise. In other embodiments, the first fixing part 1320 at the top of each support column 132 is arranged in a notch shape, and each support column 132 is arranged in a roughly "concave" shape. The second fixing parts 1330 corresponding to each first fixing part 1320 on the top plate 133 are arranged in a downward convex shape from their bottom surface.

[0056] The top plate 133 is arranged above at least one culture dish 110 and is used to support the circuit board 140. At least one limiting opening 1331 is provided on the top plate 133 for accommodating the top ends of at least one culture dish 110. Compared with the circuit board 140, the top plate 133 has a greater thickness. The top plate 133 and the circuit board 140 are fixedly connected, for example, the top plate 133 and the circuit board 140 can be threadedly connected using screws. At least one limiting opening 1331 is provided on the top plate 133 and is opposite to the positioning component 1310 of the bottom plate 131. In this embodiment, three limiting openings 1331 are provided on the top plate 133, and the shape of each limiting opening 1331 is substantially the same as the shape of the horizontal cross section of the culture dish 110, and each limiting opening 1331 is used to accommodate the top opening portion of the corresponding culture dish 110. It can be understood that, since the top plate 133 has a certain thickness, the limiting opening 1331 of the top plate 133 and the circuit board 140 together form a structure similar to a limiting groove. In other words, the top portion of the culture dish 110 enters into the limiting groove structure, so that the limiting groove structure plays a role in limiting the culture dish 110 to prevent the culture dish 110 from shifting relative to the mounting plate assembly.

[0057] The circuit board 140 is fixedly arranged above the top plate 133 and is located above at least one culture dish 110. The shape of the circuit board 140 is roughly the same as that of the top plate 133. The circuit board 140 has a plurality of notches 144 arranged corresponding to the second fixing part 1330 of the top plate 133. The circuit board 140 is provided with a plurality of sockets 142 for electrically connecting at least one group of electrode plate groups 120. In this embodiment, the at least one group of electrode plate groups 120 is welded to a side of the circuit board 140 close to the top plate 133 through the corresponding sockets 142 to achieve electrical connection. The at least one group of electrode plate groups 120 extends into the culture dish 110 in a direction away from the circuit board 140 through the corresponding opening 1101 of the culture dish 110. A group of electrode plate groups 120 includes at least one group of electrode plates 121. In this embodiment, the electrode plate groups 120 are provided with 3 groups, and the number of electrode plate groups 120 in each group is 4. There are 16 sockets 142 corresponding to each electrode plate group 120. In other embodiments, the circuit board 140 is provided with at least one group of sockets (not shown) on one side close to the top plate 133, and one end of each group of sockets (not shown) is electrically connected to the circuit board 140 through the corresponding sockets 142, and the other end is electrically connected to the corresponding electrode plate group 120. Each of the aforementioned electrode plates 121 is electrically connected to the corresponding sockets 142 or sockets (not shown) through its four conductive terminals 1213.

[0058] A cable connection terminal 143 is provided at one end of the circuit board 140. A relief notch 1332 is provided between the top plate 133 and the cable connection terminal 143 of the circuit board 140 to accommodate the cable connection terminal 143 of the circuit board 140. The cable connection terminal 143 is used to connect a cable (not shown). One end of the cable (not shown) is electrically connected to the circuit board 140 through the cable connection terminal 143, and then electrically connected to multiple electrode plates 121 through the circuit board 140; the other end is electrically connected to the adapter 420, and then electrically connected to the electric field generator 410 through the adapter 420. In other embodiments, the other end of the cable (not shown) can also be directly electrically connected to the electric field generator 410. Since the culture device 100 is placed in the incubator 200 for culture, and the electric field generator 410 and the adapter 420 are outside the incubator 200, in order not to affect the normal operation of the incubator 200, a thinner cable is selected to transmit the electrical signal between the culture device 100 and the adapter 420 or the electric field generator 410.

[0059] The circuit board 140 is also provided with at least one pinhole 141 in the area surrounded by the plurality of jacks 142, so that the corresponding temperature measuring line 320 of at least one temperature measuring line 320 of the thermometer 300 can pass through to enter the corresponding culture dish 110 of at least one culture dish 110. The pinhole 141 is approximately located at the center between the plurality of jacks 142. In this embodiment, the test instrument 330 of the thermometer 300 is arranged outside the culture device 100, and each temperature measuring line 320 of the thermometer 300 enters the corresponding culture dish 110 through the pinhole 141 of the circuit board 140, thereby allowing the thermocouple probe to contact the culture solution. Since the area surrounded by the plurality of jacks 142 corresponds to the limit opening 1331 of the top plate 133, the temperature measuring line 320 can be directly extended into the inside of the culture dish 110 without being blocked by the top plate 133.

[0060] Figure 8 The figure shows a schematic diagram of the culture device 100 during the assembly process according to an embodiment of the present disclosure. When assembling the culture device 100, the plurality of electrode plates 121 can be first welded to the circuit board 140, and then the top plate 133 and the circuit board 140 are assembled, and then the bottom plate 131 and the support column 132 are fixedly installed. Then, the culture dish 110 filled with the culture fluid is placed at the position of the positioning component 1310 on the bottom plate 131. Finally, the plurality of second fixing components 1330 of the top plate 133 are spliced ​​with the first fixing components 1320 of the corresponding support column 132. Figure 3 As shown, after the culture device 100 is assembled, the culture dish 110 enters the limiting opening 1331 , and the electrode plate 121 extends into the interior of the culture dish 110 .

[0061] The incubator 200 includes a temperature regulator 210, and the temperature regulator 210 is configured to adjust the internal temperature of the incubator 200. In the present embodiment, the incubator 200 may be a carbon dioxide incubator, and the experimental operator may operate the temperature regulator 210 to keep the internal temperature of the carbon dioxide incubator constant. In the present embodiment, the carbon dioxide incubator 200 is a water-jacketed carbon dioxide incubator, and its temperature regulator 210 is an independent water jacket layer surrounding the incubator body, and the water in the water jacket layer may be heated by an electric heating wire, and the temperature change may be detected by a temperature sensor (not shown) provided inside the box body, so that the temperature inside the box is kept constant at the set temperature. In addition, the incubator 200 may also include a carbon dioxide regulator (not shown) for adjusting the concentration of carbon dioxide inside the incubator 200, so that the concentration of carbon dioxide inside the incubator meets the requirements of the culture device 100.

[0062] The temperature measuring device 300 is used to detect the temperature of the culture solution contained in at least one culture dish 110. In some embodiments, the temperature measuring device 300 can be an instrumented temperature measuring device, such as a thermocouple temperature measuring device. The thermocouple temperature measuring device 300 includes at least one temperature measuring wire 320. The end of the temperature measuring wire 320 is a thermocouple probe (not shown). The at least one thermocouple probe is used to extend into the culture solution contained in at least one culture dish 110 to generate at least one corresponding detection signal indicating the temperature of the culture solution contained in one culture dish 110. The test meter 330 can be arranged outside the incubator 200, and is used to convert the at least one detection signal into a corresponding temperature value. At least one temperature measuring wire 320 is respectively connected to the at least one thermocouple probe (not shown) and the test meter 330, and is used to transmit the at least one detection signal to the test meter 330. The thermocouple probe (not shown) is made of alloy, has higher waterproof performance than the temperature sensor 1214 on the electrode plate 121, and has higher accuracy. Combining the thermometer 300 with the temperature sensor 1214 can provide double protection for the detection of the temperature of the culture dish 110. When the temperature sensor 1214 fails due to water seeping into the sealant (not shown), the data detected by the thermometer 300 can be used as a basis to adjust the heating temperature of the incubator 200 in time.

[0063] The alternating electric field applied by the electrode plate group 120 forms a current flowing through the culture fluid, and the cell culture fluid itself has dielectric loss, and the culture fluid generates heat. The heat generated by the culture fluid will further diffuse to the air environment inside the incubator 200. In this embodiment, in response to detecting that the temperature of the culture fluid exceeds the temperature threshold range of 36°C-38°C, the temperature regulator 210 of the incubator 200 can be operated to adjust the internal temperature of the incubator 200 so that the temperature of the culture fluid falls within the temperature threshold range of 36°C-38°C. The internal temperature inside the incubator is controlled according to the detected culture fluid temperature, thereby maintaining a balance between heat generation and heat dissipation of the entire culture fluid, so that the culture fluid is at a temperature suitable for culturing biological tissues, thereby improving the culture efficiency of the culture device 100 of the in vitro experimental device.

[0064] In some related technologies, there is a solution for controlling the output energy by adjusting the alternating voltage on the electrode plate 121 of the culture device 100, thereby stabilizing the temperature of the culture solution. However, the in vitro experimental device using this method will cause output voltage fluctuations during the electric field action, causing fluctuations in the magnitude of the field, which may be disadvantageous for the repeatability of the experiment itself in in vitro research, the quantitative statistics of the experimental results, etc. In contrast, according to this embodiment, the temperature of the culture solution is stabilized by adjusting the internal temperature of the incubator 200, thereby avoiding adjusting the voltage on the electrode plate 121, and preventing the fluctuation of the magnitude of the field from affecting the experiment.

[0065] The above-mentioned temperature threshold range of 36°C-38°C is a pre-set temperature range, which represents the temperature range required for the culture medium to culture cell tissues. The temperature range can be set according to the ideal temperature of the culture medium for culturing cell tissues. At this ideal temperature, the culture medium can achieve the best culture effect. For example, the above-mentioned ideal temperature can be 37°C, so the temperature threshold range can be set to a temperature range with a fluctuation of 1°C based on 37°C, that is, 36°C to 38°C. If it is detected that the culture medium temperature exceeds the temperature threshold range, the temperature regulator 210 of the control incubator 200 adjusts the internal temperature of the incubator 200 so that the culture medium temperature falls within the temperature threshold range again.

[0066] In some embodiments, the in vitro experimental system 1000 may also include a controller (not shown), which is configured to automatically control the temperature regulator 210 of the incubator 200 to adjust the internal temperature of the incubator so that the temperature of the culture solution falls within the temperature threshold range in response to detecting that the temperature of the culture solution exceeds the temperature threshold range. The above-mentioned adjustment of the internal temperature of the incubator can be automatically performed by the controller (not shown). In one example, the controller (not shown) can be electrically connected to the thermometer 300 and the temperature regulator 210 respectively, and the thermometer 300 generates a detection signal after detecting the temperature of the culture solution, and transmits the detection signal to the controller (not shown), and the controller (not shown) controls the temperature regulator to adjust the internal temperature of the incubator 210 according to the temperature information contained in the detection signal. In this context, the controller (not shown) includes a microcontroller or computer that executes instructions stored in firmware and / or software. The controller is programmable to perform the functions described herein. As used herein, the term computer is not limited only to integrated circuits known in the art as computers, but broadly refers to computers, processors, microcontrollers, microcomputers, programmable logic controllers, application specific integrated circuits, and other programmable circuits, and these terms are used interchangeably herein.

[0067] In some other embodiments, the above-mentioned adjustment of the internal temperature of the incubator 200 can be performed manually. In one example, the temperature detector 300 may also additionally include an alarm (not shown), which may be, for example, a sound signal alarm or a light signal alarm. The alarm is configured to generate an alarm signal when the temperature detector 300 detects that the temperature of the culture solution exceeds the temperature threshold range. After hearing / seeing the alarm signal, the relevant experimental operator can adjust the internal temperature of the incubator by manually setting the operating temperature of the temperature regulator.

[0068] The air conditioner 500 and the incubator 200 are arranged in the same space environment (for example, in the same room). Since the above-mentioned incubator 200 has no refrigeration function, and although the heat preservation ability of the incubator 200 is good, the heat conductivity is poor, so sometimes the heat generated by the culture device 100 will be greater than the heat transferred to the outside by the incubator 200, resulting in the temperature of the environment in the incubator 200 being out of control. For this reason, the ambient temperature of the in vitro experiment system 1000 can be adjusted to a low temperature. For example, the operating temperature of the air conditioner 500 can be adjusted to a temperature lower than that of the incubator 200, and the heat of the incubator 200 can be dissipated through the air environment of the in vitro experiment system 1000. The air conditioner 500 can be set to a lower operating temperature (generally lower than the temperature of the culture solution in the culture device 100) so that the culture solution in the culture device 100 is kept within a stable temperature range.

[0069] The following will refer to Fig. 9 and Fig.10 The temperature regulation method for the in vitro experimental system 1000 according to an embodiment of the present disclosure is described in detail. Fig. 9 A flow chart of a temperature regulation method 2000 for an in vitro experimental system 1000 according to an embodiment of the present disclosure is shown. The method 2000 includes:

[0070] Step 2001, using the temperature detector 300 to periodically detect the temperature of the culture solution contained in at least one culture dish 110; and

[0071] Step 2002: In response to detecting that the culture solution temperature exceeds a temperature threshold range, the temperature regulator 210 is controlled to adjust the internal temperature of the incubator 200 so that the culture solution temperature falls within the temperature threshold range.

[0072] In step 2001, the temperature of the culture solution contained in the culture dish 110 may be detected at preset intervals, for example, the temperature of the culture solution may be periodically detected at intervals of 10 seconds, 1 minute, 10 minutes, etc.

[0073] In step 2002, the temperature threshold range is a pre-set temperature range, which represents the temperature range required for the culture medium to culture cell tissues. The temperature range can be set according to the ideal temperature of the culture medium for culturing cell tissues, and at this ideal temperature, the best culture effect can be achieved. For example, the above-mentioned ideal temperature can be 37°C, so the temperature threshold range can be set to a temperature range of 1°C above and below 37°C, that is, 36°C to 38°C. If it is detected that the culture medium temperature exceeds the temperature threshold range, the temperature regulator 210 is controlled to adjust the internal temperature of the incubator 200 so that the culture medium temperature falls within the temperature threshold range again. In some embodiments, the temperature regulator 210 has the function of adjusting its operating temperature. For example, a control panel (not shown) can be provided on the incubator 200, which is connected to the temperature regulator 210 and is used to adjust the operating temperature of the temperature regulator 210. If it is detected that the culture medium temperature is higher than 38°C, the operating temperature currently set by the temperature regulator 210 can be appropriately lowered so that the internal temperature of the incubator 200 drops and falls back into the temperature threshold range. If it is detected that the culture liquid temperature is lower than 36° C., the operating temperature currently set by the temperature regulator 210 can be appropriately increased so that the internal temperature of the incubator 200 rises and falls back into the temperature threshold range.

[0074] Fig.10 A flow chart of a temperature adjustment method 3000 for an in vitro experimental system 1000 according to another embodiment of the present disclosure is shown. The temperature adjustment method 3000 can be applied to the in vitro experimental system 1000. The method 3000 includes:

[0075] Step 3001, respectively setting the initial operating temperatures of the air conditioner 500 and the temperature regulator 210;

[0076] Step 3002, using the temperature detector 300 to detect the temperature of the culture solution contained in at least one culture dish 110;

[0077] Step 3003, whether the detected culture solution temperature exceeds the upper limit threshold, if the judgment result is that the detected culture solution temperature exceeds the upper limit threshold, then execute step 3004, if the judgment result is that the detected culture solution temperature does not exceed the upper limit threshold, then execute step 3006;

[0078] Step 3004, generate an alarm signal and execute step 3005;

[0079] Step 3005, setting the target operating temperature of the temperature regulator 210 based on the difference between the current operating temperature and the culture solution temperature and the ideal temperature value, and executing step 3002;

[0080] Step 3006, whether the detected culture solution temperature exceeds the lower limit threshold, if the judgment result is that the detected culture solution temperature exceeds the lower limit threshold, then execute step 3004; if the judgment result is that the detected culture solution temperature does not exceed the lower limit threshold, then execute step 3007;

[0081] Step 3007, the temperature setting of the temperature regulator 210 is not changed, and step 3002 is repeatedly executed.

[0082] Before step 3001, firstly, the electric field generator 410 is connected to the adapter 420, the adapter 420 is connected to the culture device 100, the culture device 100 is placed in the incubator 200, and the thermocouple probe of the thermometer 300 is inserted into the culture solution. In addition, the temperature threshold range can be set to 36°C to 38°C in advance, that is, the upper alarm threshold of the alarm of the thermometer 300 is set to 38°C, and the lower alarm threshold is set to 36°C. In step 3001, the initial operating temperature of the incubator 200 can be set to 30°C, the carbon dioxide concentration can be set to 5%, and the initial operating temperature of the air conditioner 500 can be set to 24°C. After preparation, turn on the electric field generator 410, and the thermometer 300 continuously detects the temperature of the culture solution.

[0083] In step 3003, the upper threshold is 38°C.

[0084] In step 3004 , when the temperature of the cell culture fluid detected by the thermometer 300 is greater than 38° C., the alarm signal is sent by the alarm in the thermometer 300 .

[0085] In step 3005, the operating temperature of the incubator 200 is adjusted according to the following formula:

[0086] Ts=Tc-Tl+37℃

[0087] Wherein, Ts is the target operating temperature to be set for the incubator 200, Tc is the current operating temperature currently set for the incubator 200, and Tl is the temperature of the culture fluid. As an example, if the current operating temperature of the incubator 200 is 32°C, and the culture fluid temperature is detected to reach 39°C during the detection process, the alarm will send out an alarm signal, and the target operating temperature calculated by the above formula is 30°C. Therefore, the operating temperature of the temperature regulator 210 will be adjusted from the original 32°C to 30°C. After the operating temperature of the temperature regulator 210 is lowered, the temperature of the culture fluid will drop.

[0088] In step 3006, the lower threshold is 36°C.

[0089] When the culture liquid temperature detected by the thermometer 300 is less than or equal to 38° C., it is determined whether the temperature is less than 36° C. If it is not less than 36° C., the temperature setting of the temperature regulator 210 is not changed, that is, the current operating temperature of the temperature regulator 210 is maintained. Otherwise, the alarm sends an alarm signal to prompt the user to adjust the operating temperature of the incubator 200 according to the above formula.

[0090] As an example, if the current operating temperature of the incubator 200 is 32°C, and the culture solution temperature is detected to be only 35°C during the detection process, the alarm will send out an alarm signal, and the target operating temperature calculated by the above formula is 34°C. Therefore, the operating temperature of the temperature regulator 210 will be adjusted from the original 32°C to 34°C. After the operating temperature of the temperature regulator 210 is increased, the temperature of the culture solution will rise.

[0091] After step 3005 and step 3007, the above temperature detection and control of the operating temperature of the incubator 200 may be repeated until the temperature zone of the culture solution becomes stable and no longer triggers the alarm device to send out an alarm signal.

[0092] In some embodiments, the above-mentioned adjustment of the operating temperature of the temperature regulator 210 can be automatically performed by a controller. In one example, the controller can be electrically connected to the thermometer 300 and the temperature regulator 210 respectively. After the thermometer 300 detects the temperature of the culture solution, it generates a detection signal and transmits the detection signal to the controller. The controller controls the temperature regulator 210 to adjust the internal temperature of the incubator 200 according to the temperature information contained in the detection signal.

[0093] In some other embodiments, the above adjustment of the internal temperature of the incubator 200 can be performed manually. In one example, after receiving the alarm signal of the alarm, the relevant experimental operator can manually set the operating temperature of the temperature regulator 210.

[0094] The in vitro experimental system 1000 of the present invention uses a thermometer 300 to detect the temperature inside the culture dish 110. When it detects that the temperature of the culture solution exceeds the temperature threshold range, it adjusts the internal temperature of the incubator 200 by adjusting the temperature regulator 210 of the incubator 200 so that the temperature of the culture solution falls within the temperature threshold range. There is no need to adjust the alternating voltage, thereby avoiding the influence of fluctuations in the field strength on the experiment, which is beneficial to the repeatability of the experiment itself in the in vitro research, the quantitative statistics of the experimental results, etc.

[0095] The above is only a preferred implementation mode of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the scope of protection of the present application.

Claims

1. An in vitro experimental system, It is characterized in that include: an incubator, the incubator comprising a temperature regulator configured to regulate an internal temperature of the incubator; A culture device, arranged inside the culture box, comprises: at least one culture dish for containing a culture solution for culturing biological tissue; and at least one set of electrode plates, each set of electrode plates being at least partially disposed in a corresponding one of the at least one culture dish, for applying an electric field to a culture solution contained in the culture dish; a temperature detector for detecting the temperature of the culture solution contained in the at least one culture dish; and a controller, wherein the controller is configured to automatically control the temperature regulator to adjust the internal temperature of the incubator so that the temperature of the culture solution falls within the temperature threshold range in response to detecting that the temperature of the culture solution exceeds the temperature threshold range, The temperature threshold range includes an ideal temperature value that the culture solution is expected to maintain, and controlling the temperature regulator to adjust the internal temperature of the incubator includes: obtaining a current operating temperature set for the temperature regulator; and The target operating temperature of the temperature regulator is set by summing the difference between the current operating temperature and the culture solution temperature and the ideal temperature value.

2. The in vitro experimental system according to claim 1, It is characterized in that The temperature detector comprises: at least one temperature measuring line, each temperature measuring line having a thermocouple probe at the end thereof, the thermocouple probe extending into the culture liquid in the corresponding culture dish; and A test instrument is used to convert at least one detection signal into a corresponding temperature value.

3. The in vitro experimental system according to claim 2, It is characterized in that The culture device further comprises: a circuit board, which is arranged above the at least one culture dish and is used for electrically connecting at least one group of electrode plates.

4. The in vitro experimental system according to claim 3, It is characterized in that The circuit board is provided with a plurality of sockets for plugging in the at least one group of electrode plates, and at least one pinhole for the temperature measuring line to pass through is also provided in the area surrounded by the plurality of sockets. The at least one culture dish is provided with a corresponding opening at its respective top end. The at least one group of electrode plates is arranged on the circuit board through the plurality of sockets, and extends into the at least one culture dish through the corresponding opening of the at least one culture dish in a direction away from the circuit board.

5. The in vitro experimental system according to claim 4, It is characterized in that Each electrode plate group includes at least two pairs of electrode plates.

6. The in vitro experimental system according to claim 4, It is characterized in that Each electrode plate includes a flexible circuit board, an insulating substrate and a ceramic electrode arranged on two opposite sides of the flexible circuit board.

7. The in vitro experimental system according to claim 6, It is characterized in that The flexible circuit board has the same shape as the insulating substrate, and both are arranged in a "T" shape.

8. The in vitro experimental system according to claim 7, It is characterized in that The width of the flexible circuit board at one end close to the circuit board is smaller than the width of the flexible circuit board at one end away from the circuit board.

9. The in vitro experimental system according to claim 6, It is characterized in that The ceramic electrode is arranged at one end of the flexible circuit board away from the circuit plug board.

10. The in vitro experimental system according to claim 6, It is characterized in that Each electrode plate also includes at least one temperature sensor arranged at one end of the flexible circuit board away from the circuit plug board, and the temperature sensor is arranged on the same side of the flexible circuit board as the ceramic electrode.

11. The in vitro experimental system according to claim 10, It is characterized in that Each electrode plate also includes a plurality of conductive terminals arranged at one end of the flexible circuit board close to the circuit board, one end of each conductive terminal is electrically connected to the flexible circuit board, and the other end is electrically connected to the circuit board above the culture dish.

12. The in vitro experimental system according to claim 11, It is characterized in that The other end of each conductive terminal is electrically connected to a corresponding plug hole on the circuit board.

13. The in vitro experimental system according to claim 3, It is characterized in that The culture device further comprises: a mounting plate assembly for mounting and fixing the at least one culture dish and the circuit board.

14. The in vitro experimental system according to claim 13, It is characterized in that The mounting plate assembly comprises a bottom plate, a plurality of supporting columns fixed on the bottom plate, and a top plate detachably fixed to the plurality of supporting columns.

15. The in vitro experimental system according to claim 14, It is characterized in that The bottom plate is a plate-shaped structure and is used to support at least one culture dish.

16. The in vitro experimental system according to claim 15, It is characterized in that A side surface of the bottom plate supporting the culture dish is provided with at least one positioning component for limiting the horizontal movement of the culture dish.

17. The in vitro experimental system according to claim 14, It is characterized in that A first fixing component is provided on the top of each supporting column.

18. The in vitro experimental system according to claim 17, It is characterized in that The top plate is provided with a plurality of second fixing parts at positions corresponding to the first fixing parts of the plurality of support columns in a one-to-one correspondence.

19. The in vitro experimental system according to claim 18, It is characterized in that The top plate is detachably fixedly connected to the first fixing parts of the corresponding support columns through its multiple second fixing parts.

20. The in vitro experimental system according to claim 14, It is characterized in that The top plate is arranged above at least one culture dish and is used for supporting the circuit board.

21. The in vitro experimental system according to claim 20, It is characterized in that At least one limiting opening is formed on the top plate, for accommodating the top ends of at least one culture dish respectively.

22. The in vitro experimental system according to claim 3, It is characterized in that A cable connection terminal is provided at one end of the circuit board, and the cable connection terminal is used to connect the cable.

23. The in vitro experimental system according to any one of claims 1 to 22, It is characterized in that Also includes: The electric field generator is used to generate at least one alternating electric field signal and transmit it to at least one group of electrode plates.

24. The in vitro experimental system according to claim 23, It is characterized in that It also includes an adapter, which is electrically connected to the electric field generator and is used to provide the at least one alternating electric field signal to at least one group of electrode plates through the adapter.

25. The in vitro experimental system according to claim 1, It is characterized in that The temperature threshold range is 36°C-38°C.

26. A method for regulating the temperature of an in vitro experimental system, wherein the in vitro experimental system is the in vitro experimental system according to any one of claims 1 to 25, It is characterized in that The method comprises: using the temperature detector to periodically detect the temperature of the culture solution contained in the at least one culture dish; and In response to detecting that the culture solution temperature exceeds a temperature threshold range, controlling the temperature regulator to adjust the internal temperature of the incubator so that the culture solution temperature falls within the temperature threshold range, wherein the temperature threshold range is 36° C.-38° C., and the temperature threshold range includes an ideal temperature value that the culture solution is expected to maintain. Wherein, controlling the temperature regulator to adjust the internal temperature of the incubator further comprises: obtaining a current operating temperature set for the temperature regulator; and The target operating temperature of the temperature regulator is set by summing the difference between the current operating temperature and the culture solution temperature and the ideal temperature value.

27. The temperature adjustment method according to claim 26, It is characterized in that Also includes: In response to detecting that the culture fluid temperature does not exceed the temperature threshold range, the temperature setting of the temperature regulator is not changed.

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