CN114149922A

CN114149922A - In-vitro experiment system and temperature adjusting method thereof

Info

Publication number: CN114149922A
Application number: CN202111599549.3A
Authority: CN
Inventors: 惠嘉杰; 陈晟; 冯晶晶
Original assignee: Jiangsu Hailai Xinchuang Medical Technology Co Ltd
Current assignee: Jiangsu Hailai Xinchuang Medical Technology Co Ltd
Priority date: 2021-12-24
Filing date: 2021-12-24
Publication date: 2022-03-08

Abstract

The invention provides an in vitro experiment system, which comprises: an incubator including a temperature regulator that regulates an internal temperature of the incubator; culture apparatus sets up inside the incubator, includes: at least one culture dish for containing a culture solution for culturing biological tissue, and at least one set of electrode plates for applying an electric field to the culture solution contained in the respective culture dish; and a temperature detector for detecting a temperature of the culture solution contained in the at least one culture dish, wherein the temperature regulator is configured to regulate an internal temperature of the incubator such that the temperature of the culture solution falls within a temperature threshold range when the temperature detector detects that the temperature of the culture solution is outside the temperature threshold range. The in-vitro experiment system provided by the invention adjusts the internal temperature of the incubator by adjusting the temperature adjuster of the incubator so that the temperature of the culture solution falls within the temperature threshold range, and the influence of the fluctuation of the field intensity on the experiment is avoided without adjusting the alternating voltage.

Description

In-vitro experiment system and temperature adjusting method thereof

Technical Field

The invention relates to the field of in-vitro experimental equipment, in particular to an in-vitro experimental system and a temperature adjusting method thereof.

Background

Tumor therapeutic electric fields (TTFields) are a therapeutic approach to inhibit tumor cell proliferation by interfering with tumor cell mitosis. The low-intensity medium-high frequency alternating electric field is used for intervening and treating various tumor diseases, and is a clinically effective cancer treatment method. Many basic studies have been conducted at home and abroad to confirm the biological effects of TTfields. Cell experiment research results show that TTfields can regulate and control biochemical molecule expression in tumor cells, interfere division cycles of the tumor cells, inhibit proliferation of the tumor cells and promote apoptosis of the tumor cells. Animal experiment research results show that TTfields can effectively inhibit the tumor angiogenesis in tumor-bearing mice and inhibit the growth of tumors in the mice. The current results of clinical research at home and abroad also prove the positive effect of TTfields on tumor treatment.

In vitro studies, TTFields act on the cell culture fluid in a cell culture device to generate a large amount of thermal energy, which causes the temperature of the culture fluid to rise continuously. However, the requirement of the in vitro culture of the biological tissue on the environmental temperature is high, the cell culture solution needs to be maintained at about 37 ℃, otherwise, the biological tissue is easy to die due to inappropriate environmental temperature, and the referential of the TTfields in vitro experiment is affected. The temperature of the culture solution disclosed in the prior patent publication No. 112680349 is monitored by a heat-sensitive sensor disposed on the electrode plate, a temperature signal monitored by the heat-sensitive sensor is transmitted to an external control unit, and the external control unit adjusts the output intensity of the AC signal according to the temperature signal to control the heat generated by the electric field, so as to maintain the temperature of the cell culture solution at about 37 ℃. The above-mentioned method for maintaining the temperature of the cell culture fluid is realized by adjusting the output intensity of the electric signal, which causes the fluctuation of the field intensity of the alternating electric field, and the fluctuation field intensity is disadvantageous to the repeatability of the in vitro research experiment, the quantitative statistics of the experimental result, etc. Meanwhile, the thermosensitive sensor on the electrode plate is immersed in the culture solution for a long time, so that the cell culture solution easily permeates into the sealing glue coated on the periphery of the thermosensitive sensor to influence the thermosensitive sensor, the temperature monitored by the thermosensitive sensor has errors, and the temperature of the culture solution is further influenced.

Therefore, there is a need to provide an improved in vitro experiment system and a temperature adjustment method thereof, so as to avoid the disadvantages of the in vitro experiment system and the temperature adjustment method.

Disclosure of Invention

The invention provides an in-vitro experiment system with stable field intensity and accurate temperature monitoring and a temperature adjusting method thereof.

The in vitro experiment system is realized by the following technical scheme: an in vitro assay system comprising: an incubator comprising a temperature regulator configured to regulate an internal temperature of the incubator; culture apparatus, set up inside the incubator, include: 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 fluid contained in the culture dish; and a temperature detector for detecting a temperature of the culture solution contained in the at least one culture dish, wherein the temperature regulator regulates an internal temperature of the incubator so that the culture solution temperature falls within a temperature threshold range when the temperature detector detects that the culture solution temperature is outside the temperature threshold range.

Further, the temperature detector includes: the tail end of each temperature measuring line is provided with a thermocouple probe which extends into the culture solution in the corresponding culture dish; and the test instrument is used for converting the at least one detection signal into a corresponding temperature value.

Further, the culture apparatus further comprises: and the circuit plugboard is arranged above the at least one culture dish and is used for electrically connecting at least one group of electrode plates.

Further, be provided with on the circuit insertion board and be used for pegging graft a plurality of jacks of at least a set of electrode plate still are equipped with at least one pinhole that is used for the temperature measurement line to pass in the region that a plurality of jacks encircle, at least one culture dish is provided with corresponding opening in respective top department, at least a set of electrode plate passes through a plurality of jacks set up on the circuit insertion board, and pass through corresponding opening of at least one culture dish is along keeping away from the direction of circuit insertion board extends to in the at least one culture dish.

Further, each group of electrode plate groups at least comprises two pairs of electrode plates.

Furthermore, each electrode plate comprises a flexible circuit board, an insulating substrate and a ceramic electrode, wherein the insulating substrate and the ceramic electrode are arranged on two opposite sides of the flexible circuit board.

Furthermore, the flexible circuit board and the insulating substrate are approximately same in shape and are both approximately arranged in a T shape.

Furthermore, the width of one end of the flexible circuit board close to the circuit board is smaller, and the width of one end of the flexible circuit board far away from the circuit board is larger.

Furthermore, the ceramic electrode is arranged at one end of the flexible circuit board far away from the circuit plug board.

Furthermore, each electrode plate further comprises at least one temperature sensor arranged at one end, far away from the circuit board, of the flexible circuit board, and the temperature sensors are arranged on the flexible circuit board and located on the same side with the ceramic electrodes.

Furthermore, each electrode plate further comprises a plurality of conductive terminals arranged at one end, close to the circuit board, of the flexible circuit board, one end of each conductive terminal is electrically connected with the flexible circuit board, and the other end of each conductive terminal is electrically connected with the circuit board above the culture dish.

Furthermore, the other end of each conductive terminal is electrically connected with a corresponding jack on the circuit plugboard respectively.

Further, the culture apparatus further comprises: and the mounting plate assembly is used for mounting and fixing the at least one culture dish and the circuit insertion plate.

Furthermore, the mounting plate assembly comprises a bottom plate, a plurality of supporting columns fixed on the bottom plate and a top plate detachably fixed with the supporting columns.

Further, the base plate is of generally plate-like configuration for supporting at least one culture dish.

Further, the bottom plate supports at least one positioning part for limiting the culture dish to move horizontally on one side surface of the culture dish.

Further, the top of each support column is provided with a first fixing part.

Furthermore, a plurality of second fixing parts are arranged on the top plate at positions corresponding to the first fixing parts of the support columns in a one-to-one correspondence manner.

Further, the top plate is detachably fixedly connected with the first fixing parts of the corresponding support columns through a plurality of second fixing parts of the top plate.

Further, the top plate is arranged above the at least one culture dish and is used for supporting the circuit board.

Furthermore, the top plate is provided with at least one limiting opening for respectively accommodating the top ends of at least one culture dish.

Furthermore, a flat cable connecting terminal is arranged at one end of the circuit plug board and used for connecting a flat cable.

Further, the method also comprises the following steps: and the electric field generator is used for generating at least one alternating electric field signal and transmitting the alternating electric field signal to at least one group of electrode plates.

Further, the device also comprises an adapter, wherein the adapter is electrically connected with the electric field generator and is used for providing the at least one alternating electric field signal to at least one group of electrode plates through the adapter.

Further, the method also comprises the following steps: a controller configured to automatically control the temperature regulator to adjust the internal temperature of the incubator such that the culture fluid temperature falls within a temperature threshold range in response to detecting that the culture fluid temperature is outside the temperature threshold range.

Further, the temperature threshold range is 36-38 ℃.

The temperature regulation method of the in-vitro experiment system is realized by the following scheme: a method of temperature regulation in an in vitro assay system as described above, the method comprising: periodically detecting the temperature of the culture solution contained in the at least one culture dish by using the temperature detector; and in response to detecting that the culture fluid temperature is outside a temperature threshold range, controlling the temperature regulator to regulate the internal temperature of the incubator such that the culture fluid temperature falls within the temperature threshold range, the temperature threshold range being 36-38 ℃.

Further, the temperature threshold range includes a desired temperature value at which the culture solution is expected to be maintained, wherein controlling the temperature regulator to regulate the internal temperature of the incubator further includes: acquiring a current operating temperature set for the temperature regulator; and setting a target operating temperature of the thermostat based on a difference between the current operating temperature and the culture solution temperature and the desired temperature value.

Further, the method also comprises the following steps: in response to detecting that the culture fluid temperature does not exceed the temperature threshold range, not changing the temperature setting of the thermostat.

The in-vitro experiment system provided by the invention utilizes the temperature measurer to detect the temperature in the culture dish, when the temperature of the culture medium is detected to exceed the temperature threshold range, the temperature inside the culture box is adjusted by adjusting the temperature adjuster of the culture box, so that the temperature of the culture medium is within the temperature threshold range, the alternating voltage is not required to be adjusted, the influence of the fluctuation of the field intensity on the experiment is avoided, and the repeatability of the experiment in-vitro research, the quantitative statistics of the experiment result and the like are facilitated.

These and other aspects of the disclosure will be apparent from and elucidated with reference to the embodiments described hereinafter.

Drawings

FIG. 1 is a schematic block diagram of an in vitro experimental system according to the present invention.

FIG. 2 is a perspective view of the culture apparatus and the temperature detector in FIG. 1;

FIG. 3 is a schematic view of the section A-A of the culture apparatus in FIG. 2;

FIG. 4 is an exploded perspective view of the culture device of FIG. 2;

FIG. 5 is a schematic plan view of an electrode plate of the culture apparatus in FIG. 4; and is

FIG. 6 is an exploded perspective view of the electrode plate shown in FIG. 5;

FIG. 7 is a perspective view of a circuit board of the culture device in FIG. 4;

FIG. 8 is a schematic view of the culture device of FIG. 2 during assembly;

FIG. 9 is a flowchart of a temperature adjustment method for an in vitro assay system according to a first embodiment of the present invention;

FIG. 10 is a flowchart of a temperature adjustment method for an in vitro experiment system according to a second embodiment of the present invention.

Detailed Description

Reference will now be made in detail to the exemplary embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, like numbers in different drawings represent the same or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present application. Rather, they are merely examples of electrode sheets consistent with certain aspects of the present application as detailed in the appended claims.

Fig. 1 to 8 show an in vitro experiment system 1000 according to the present invention, which includes a culture apparatus 100, an incubator 200 in which the culture apparatus 100 is placed, a thermocouple-type temperature detector 300 for monitoring the temperature of a culture fluid in the culture apparatus 100, an electric field generator 410 for supplying an alternating electric signal, an adaptor 420 for transmitting the alternating electric signal from the electric field generator 410 to the culture apparatus, and an air conditioner 500 for controlling the ambient temperature (room temperature) of a space of the in vitro experiment system 1. The incubator 200 includes a temperature regulator 210 that regulates a heating temperature of the incubator 200 to regulate an ambient temperature inside the incubator 200. The in vitro experiment 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 experiment system 1000 adjusts the heating temperature of the temperature regulator 210 according to the temperature of the culture solution monitored by the temperature detector 300, so that the temperature of the culture solution in the culture device 100 placed in the incubator 200 falls within the temperature threshold range of 36-38 ℃, and the air conditioner 500 is used for assisting in adjusting the environmental temperature in the incubator 200, so that the heat in the incubator 200 is transferred to the space environment outside the incubator 200, thereby ensuring that the cell culture solution performs TTF in vitro research experiments under the field intensity environment of a stable alternating electric field, and being beneficial to the repeatability of the in vitro research experiments and the quantitative statistics of experimental results. In some cases, the air conditioner 500 in the in vitro testing system 1000 is not necessary, for example, the temperature of the space environment is lower than the temperature in the incubator 200, and 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 experiment system 1000 is necessary, for example, the temperature of the space environment is higher than the temperature in the incubator 200, the heat in the incubator 200 cannot be transferred to the space environment outside the incubator 200, the temperature of the space environment outside the incubator 200 needs to be reduced to be lower than the temperature in the incubator 200 by the air conditioner 500, and the heat in the incubator 200 is transferred to the space environment outside the incubator 200.

The culture device 100 comprises at least one culture dish 110, at least one group of electrode plate groups 120, a mounting plate group 130 for fixing the at least one culture dish 110, and a circuit board 140 positioned above the at least one culture dish 110 and electrically connected with the at least one group of electrode plate groups 120. The culture dish 110 contains a culture solution for culturing biological tissues. The electrode plate set 120 is at least partially disposed within the culture dish 110 for applying an electric field to the culture fluid contained in the culture dish 110. In this embodiment, the number of the culture dish 110 and the electrode plate group 120 may be 3, so as to constitute different control groups. Each set of electrode plate groups 120 is disposed in a corresponding one of the culture dishes 110. The 3 culture dishes 110 are arranged at intervals along the linear direction. The size and shape of each culture dish 110 are the same. In this embodiment, each culture dish 110 is of a cubic structure, and the culture dish 110 is provided at a top end thereof with a corresponding opening 1101, through which opening 1101 culture liquid can be fed into the interior of the culture dish 110. In other embodiments, the number of the culture dishes 110 may be more than 3 or less than 3, and the culture dishes 110 may have other shapes than a cube, such as a rectangular parallelepiped, a cylindrical structure, etc., and other arrangements besides a linear arrangement, which are not listed here.

Each set of electrode plates 120 may include 4 electrode plates 121, and each pair of electrode plates 121 includes two electrode plates 121 facing each other and having opposite polarities for applying a uniform electric field to the culture solution, and the 4 electrode plates 121 constitute two pairs of electrode plates 121. In other embodiments, each electrode plate group 120 may further include other numbers of electrode plates 121, and each electrode plate group 120 may also be disposed in other suitable arrangements, which are not listed here.

Each electrode plate 121 includes a flexible circuit board 1210, an insulating substrate 1211 disposed on opposite sides of the flexible circuit board 1210, and a ceramic electrode 1212. The insulating substrate serves 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 provided in a substantially "T" shape. The end of the flexible circuit board 1210 close to the circuit board 140 has a smaller width, and the end far away from the circuit board 140 has a larger width, so as to dispose the ceramic electrode 1212 at the end with the larger width. At least one ceramic electrode 1212 of each electrode plate 121 is provided. In this embodiment, the number of the ceramic electrodes 1212 of each electrode plate 121 is two. The flexible circuit board 1210 is used for transmitting electrical signals. At least one ceramic electrode 1212 is electrically connected to the flexible circuit board 1210 to obtain an electrical signal from the flexible circuit board 1210 that applies an alternating electric field. In this embodiment, the ceramic electrode 1212 is disposed in a circular plate shape. The two ceramic electrodes 1212 are soldered to the flexible circuit board 1210 by solder (not shown) to electrically connect the two electrodes. A gap (not shown) is formed between the ceramic electrode 1212 and the flexible circuit board 1210 by soldering. A sealant (not shown) is filled in the gap (not shown), so that the ceramic electrode 1212 is prevented from being influenced by external force to break the welding part, and the alternating electric field cannot be applied to cells in the culture solution through the ceramic electrode 1212; meanwhile, it is avoided that moisture in the air enters a gap (not shown) to erode 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.

Each electrode plate 121 is further provided with at least one temperature sensor 1214. The temperature sensor 1214 is disposed 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 so as to detect the temperature of the culture solution. The temperature sensor 1214 is peripherally covered with a sealant (not shown) to protect the temperature sensor 1214. Each electrode plate 121 further includes a set of conductive terminals 1213. The set of conductive terminals 1213 is disposed on an end of the flexible circuit board 1210 proximate to the circuit card 140. One end of the set of conductive terminals 1213 is electrically connected to the flexible printed circuit 1210, and the other end is electrically connected to the circuit board 140 above the culture dish 110. The set of conductive terminals 1213 are electrically connected to the ceramic electrode 1212 and the temperature sensor 1214 respectively via the flexible circuit board 1210. In this embodiment, the number of the temperature sensors 1214 provided on each electrode plate 121 is 2. The number of the conductive terminals 1213 of each electrode plate 121 is 4.

The mounting plate assembly 130 of the culture device 100 is used for mounting and fixing at least one culture dish 110. The mounting plate assembly 130 includes a bottom plate 131, a plurality of supporting pillars 132 fixed on the bottom plate 131, and a top plate 133 detachably fixed to the plurality of supporting pillars 132.

The bottom plate 131 has a substantially rectangular plate-like configuration. The bottom plate 131 is used to support at least one culture dish 110. The bottom plate 131 is provided with at least one positioning member 1310 for limiting the horizontal movement of the culture dish 110 on the surface of the side supporting the culture dish 110. In this embodiment, there are 3 positioning components 1310. The positioning member 1310 is a groove, and the recessed area is substantially the same as the bottom surface of the culture dish 110 in shape and size. In other embodiments, the positioning member 1310 may have other shapes with the same positioning function, such as a convex ring, a plurality of convex points arranged at intervals in a ring shape, and the like.

The bottom ends of the plurality of supporting columns 132 are all fixed on the bottom plate 131, for example, the bottom end of each supporting column 132 can be fixed on the upper surface of the bottom plate 131 by a screw. The supporting columns 132 are respectively arranged between two adjacent culture dishes 110 at intervals and are positioned at the edges of two opposite sides of the bottom plate 131, so that the supporting columns 132 are avoided from the culture dishes 110, and the overall structure of the culture device 100 is more compact. The top of each support column 132 is provided with a first fixing member 1320. Each of the first fixing members 1320 is disposed in a convex shape on the top of the corresponding supporting pillar 132, and each of the supporting pillars 132 is disposed in a substantially "convex" shape. The top plate 133 is provided with a plurality of second fixing members 1330 at positions corresponding to the first fixing members 1320 of the plurality of supporting pillars 132 in a one-to-one correspondence. The second fixing members 1330 are all provided in a notch shape. The top plate 133 is detachably and fixedly connected to the first fixing parts 1320 of the corresponding support pillars 132 through a plurality of second fixing parts 1330 thereof. The second fixing parts 1330 of the top plate 133 and the first fixing parts 1320 of the corresponding support pillars 132 form a mortise and tenon joint fixing. That is, the first fixing member 1320 is a tenon, and the second fixing member 1330 is a mortise. In other embodiments, the first fixing member 1320 on the top of each supporting column 132 is disposed in a notch shape, and each supporting column 132 is disposed in a substantially concave shape. The second fixing part 1330 of the top plate 133 corresponding to each of the first fixing parts 1320 is formed to protrude downward from the bottom surface thereof.

The top plate 133 is disposed above the at least one culture dish 110 for supporting the circuit board 140, and the top plate 133 is formed with at least one position limiting opening 1331 for respectively receiving the top end of the at least one culture dish 110. The top plate 133 has a greater thickness than the circuit board 140. The top board 133 and the circuit board 140 are fixedly connected, for example, screws may be used to screw the top board 133 and the circuit board 140. The top plate 133 is provided with at least one position limiting opening 1331 opposite to the positioning part 1310 of the bottom plate 131. In this embodiment, the top plate 133 is provided with 3 limiting openings 1331, each limiting opening 1331 has a shape substantially the same as the horizontal cross section of the culture dish 110, and each limiting opening 1331 is used for accommodating the top opening portion of the corresponding culture dish 110. It can be understood that the limiting opening 1331 of the top plate 133 and the circuit board 140 form a limiting groove-like structure together because the top plate 133 has a certain thickness. That is, the top end portion of the culture dish 110 enters the limiting groove structure, so that the limiting groove structure can limit the culture dish 110 to prevent the culture dish 110 from being displaced relative to the mounting plate assembly.

The circuit board 140 is fixedly disposed above the top plate 133 and above at least one culture dish 110. The circuit board insert 140 has a shape substantially identical to the shape of the top plate 133. The circuit board 140 has a plurality of notches 144 formed corresponding to the second fixing portion 1330 of the top plate 133. The circuit board 140 is provided with a plurality of insertion holes 142 for electrically connecting at least one set of electrode plate groups 120. In this embodiment, the at least one electrode plate group 120 is electrically connected to the circuit board 140 by welding through the corresponding insertion hole 142. The at least one set of electrode plate groups 120 extends into the culture dish 110 through the corresponding opening 1101 of the culture dish 110 in a direction away from the circuit card 140. The set of electrode plates 120 includes at least one set of electrode plates 121. In this embodiment, there are 3 electrode plate groups 120, and the number of each electrode plate group 120 is 4. There are 16 insertion holes 142 corresponding to each electrode plate group 120. In other embodiments, at least one set of sockets (not shown) is disposed on the circuit board 140 near the top plate 133, one end of each set of sockets (not shown) is electrically connected to the circuit board 140 through the corresponding insertion hole 142, and the other end is connected to the corresponding electrode plate set 120. Each of the electrode plates 121 is electrically connected to the corresponding insertion hole 142 or the corresponding socket (not shown) through its four conductive terminals 1213.

The circuit board 140 has a row of connecting terminals 143 at one end. An escape notch 1332 is formed at the position of the flat cable connecting terminal 143 of the top plate 131 and the circuit board 140 to accommodate the flat cable connecting terminal 143 of the circuit board 140. The bus bar connection terminal 143 is used to connect a bus bar (not shown). One end of the flat cable (not shown) is electrically connected to the circuit plug board 140 through the flat cable connecting terminal 143, and is electrically connected to the plurality of electrode plates 121 through the circuit plug board 140; the other end is electrically connected to the adaptor 420, and then electrically connected to the electric field generator 410 through the adaptor 420. In other embodiments, the other end of the flat cable (not shown) may also be directly electrically connected to the electric field generator 410. Since the culture device 100 is placed in the incubator 200 for culturing and the electric field generator 410 and the adaptor 420 are outside the incubator 200, a thinner wire is selected between the culture device 100 and the adaptor 420 or the electric field generator 410 for conducting an electric signal in order not to affect the normal operation of the incubator 200.

The circuit board 140 further has at least one pin hole 141 in a region surrounded by the plurality of insertion holes 142 for a corresponding one 320 of the at least one temperature measuring lines 320 of the temperature detector 300 to pass through to enter the inside of a corresponding one 110 of the at least one culture dish 110. The pin hole 141 is located approximately at the center between the plurality of insertion holes 142. In this embodiment, the test meter 330 of the temperature detector 300 is disposed outside the culture apparatus 100, and each temperature measuring line 320 of the temperature detector 300 enters the corresponding culture dish 110 through the pin hole 141 of the circuit board 140, thereby allowing the thermocouple probe to contact the culture solution. Since the area surrounded by the plurality of insertion holes 142 corresponds to the position-limiting opening 1331 of the top plate 133, the temperature measuring wire 320 may not be blocked by the top plate 133 and directly extend into the culture dish 110.

FIG. 8 shows a schematic view of a culture device 100 during assembly according to one embodiment of the present disclosure. In assembling the culture apparatus 100, the plurality of electrode plates 121 may be welded to the circuit board 140, the top plate 133 and the circuit board 140 may be assembled, and the bottom plate 131 and the supporting posts 132 may be fixedly mounted. The culture dish 110 containing the culture solution is then placed on the bottom plate 131 at the position of the positioning part 1310. Finally, the plurality of second fixing members 1330 of the top plate 133 are coupled to the first fixing members 1320 of the corresponding support columns 132. As shown in FIG. 3, after the culture apparatus 100 is assembled, the culture dish 110 enters the position limiting opening 1331, and the electrode plate 121 protrudes into the culture dish 110.

Incubator 200 includes a temperature regulator 210, temperature regulator 210 configured to regulate the internal temperature of incubator 200. In this embodiment, the incubator 200 may be a carbon dioxide incubator, and the experiment operator may keep the internal temperature of the carbon dioxide incubator constant by operating the temperature regulator 210. In this embodiment, the carbon dioxide incubator 200 is a water jacket type carbon dioxide incubator, and the temperature regulator 210 is an independent water jacket layer surrounding the incubator body, and can heat water in the water jacket layer by an electric heating wire, and detect a temperature change by a temperature sensor (not shown) provided inside the incubator body, so that the temperature in the incubator is kept constant at a set temperature. In addition, the incubator 200 may further include a carbon dioxide regulator (not shown) for regulating the concentration of carbon dioxide inside the incubator 200 so that the concentration of carbon dioxide inside the incubator satisfies the demand of the culture apparatus 100.

The temperature detector 300 is used to detect the temperature of the culture solution contained in at least one culture dish 110. In some embodiments, the temperature detector 300 may be an instrumented temperature measuring device, such as a thermocouple temperature detector. The thermocouple thermometer 300 includes at least one temperature sensing wire 320. The temperature sensing wire 320 terminates in a thermocouple probe (not shown). The at least one thermocouple probe is adapted to extend into the culture fluid contained in each of the at least one culture dish 110 to generate a corresponding at least one detection signal indicative of a temperature of the culture fluid contained in one of the culture dishes 110. The test meter 330 may be disposed outside the incubator 200 for converting the at least one detection signal into a corresponding temperature value. At least one temperature measuring line 320 is connected to the at least one thermocouple probe (not shown) and the test meter 330, respectively, for transmitting the at least one detection signal to the test meter 330, respectively. The thermocouple probe (not shown) is made of alloy, and has higher waterproof performance and higher accuracy than the temperature sensor 1214 on the electrode plate 121. Combine thermoscope 300 and temperature sensor 1214, can provide dual guarantee to the detection of culture dish 110 temperature, when temperature sensor 1214 became invalid because of water infiltration sealed glue (not shown), can be accurate for the data that thermoscope 300 detected, in time adjust the temperature of heating of incubator 200.

The alternating electric field applied by the electrode plate set 120 forms a current flowing through the culture solution, and the cell culture solution itself has dielectric loss, and the culture solution generates heat. The heat generated from the culture solution is further diffused to the air atmosphere inside the incubator 200. In this embodiment, in response to detecting that the temperature of the culture liquid is outside the temperature threshold range of 36 ℃ -38 ℃, the temperature regulator 210 of the incubator 200 can be operated to regulate the internal temperature of the incubator 200 so that the temperature of the culture liquid falls within the temperature threshold range of 36 ℃ -38 ℃. The internal temperature inside the incubator is controlled according to the detected temperature of the culture solution, so that the heat production and heat dissipation balance of the whole culture solution is maintained, the culture solution is at a temperature suitable for culturing biological tissues, and the culture efficiency of the culture device 100 of the in-vitro experimental device is improved.

In some related arts, there is a scheme of stabilizing the temperature of the culture solution by controlling the output energy by adjusting the alternating voltage on the electrode plate 121 of the culture apparatus 100. However, the in vitro experimental device using this method may cause output voltage fluctuation during the electric field action process, causing the fluctuation of field intensity, which may be disadvantageous for the repeatability of the experiment itself in the in vitro research, the quantitative statistics of the experimental results, etc. Compared with the above, according to this embodiment, the stability of culture solution temperature is realized through the inside temperature of adjusting incubator 200 to avoid adjusting the voltage on electrode board 121, prevented the influence of the fluctuation of field intensity size to the experiment.

The temperature threshold range 36 ℃ to 38 ℃ is a preset temperature range that represents a temperature range required for the culture solution to culture the cell tissue. The temperature range can be set according to an ideal temperature for culturing the cell tissue by the culture solution, and the culture solution can achieve the optimal culture effect at the ideal temperature. For example, the above-mentioned ideal temperature may be 37 ℃, and then the temperature threshold range may be set to a temperature range that fluctuates by 1 ℃ from top to bottom on the basis of 37 ℃, i.e., 36 ℃ to 38 ℃. If it is detected that the culture liquid temperature is outside the temperature threshold range, the temperature regulator 210 that controls the incubator 200 regulates the internal temperature of the incubator 200 so that the culture liquid temperature falls within the temperature threshold range again.

In some embodiments, in vitro experimental system 1000 may further include a controller (not shown) configured to automatically control temperature regulator 210 of incubator 200 to regulate the internal temperature of the incubator such that the culture fluid temperature falls within a temperature threshold range in response to detecting the culture fluid temperature outside of the temperature threshold range. The adjustment of the internal temperature of the incubator can be automatically performed by a controller (not shown). In one example, a controller (not shown) may be electrically connected to the temperature detector 300 and the temperature regulator 210, respectively, and the temperature detector 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 regulate 210 the internal temperature of the incubator according to temperature information included in the detection signal. In this context, a 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 to just those integrated circuits referred to in the art as a computer, but broadly refers to a computer, a processor, a microcontroller, a microcomputer, a programmable logic controller, an application specific integrated circuit, and other programmable circuits, and these terms are used interchangeably herein.

In other embodiments, the adjustment of the internal temperature of the incubator 200 described above may be performed manually. In one example, the temperature detector 300 may additionally include an alarm (not shown), which may be, for example, an audible alarm or a light alarm. The alarm is configured to generate an alarm signal when the temperature detector 300 detects that the temperature of the culture fluid is outside of a temperature threshold range. After hearing/seeing the alarm signal, the relevant experiment operator can adjust the internal temperature of the incubator by manually setting the operating temperature of the temperature regulator.

The air conditioner 500 and the incubator 200 are disposed in the same spatial environment (e.g., the same room). Since the above-mentioned body culture tank 200 does not have a cooling function, and the heat-insulating ability of the culture tank 200 is good, but the heat-conducting ability is poor, the heat generated by the culture apparatus 100 may be larger than the heat transferred from the outside of the culture tank 200, which may cause the environmental temperature in the culture tank 200 to be out of control. To this end, the ambient temperature of the in vitro experimental system 1000 may be adjusted to a low temperature. For example, the operating temperature of the air conditioner 500 may be adjusted to be lower than the temperature of the incubator 200, and the heat of the incubator 200 may be dissipated through the air environment of the in vitro experimental system 1000. The air conditioner 500 may be set to a low operation 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 maintained in a stable temperature range.

A temperature adjustment method for the in-vitro experiment system 1000 according to an embodiment of the present disclosure will be described in detail below with reference to fig. 9 and 10. Fig. 9 shows a flow diagram of a temperature adjustment method 2000 for an in vitro experimental system 1000 according to one embodiment of the present disclosure. The method 2000 includes:

step 2001, periodically detecting the temperature of the culture solution contained in at least one culture dish 110 using a temperature detector 300; and

in response to detecting that the culture liquid temperature exceeds the temperature threshold range, the temperature regulator 210 is controlled to regulate the internal temperature of the incubator 200 so that the culture liquid temperature falls within the temperature threshold range, step 2002.

In step 2001, the temperature of the culture liquid contained in the culture dish 110 may be detected at intervals of a preset time, for example, the temperature of the culture liquid may be periodically detected at intervals of 10s, 1min, 10min, or the like.

In step 2002, the temperature threshold range is a preset temperature range that indicates a temperature range required for the culture solution to culture the cell tissue. The temperature range can be set according to an ideal temperature for culturing a cell tissue in a culture medium, and an optimal culture effect can be achieved at the ideal temperature. For example, the desired temperature may be 37 ℃, and then the temperature threshold range may be set to a temperature range that fluctuates about 37 ℃ by 1 ℃, i.e., 36 ℃ to 38 ℃. If it is detected that the culture liquid temperature is outside the temperature threshold range, the temperature regulator 210 is controlled to regulate the internal temperature of the incubator 200 so that the culture liquid temperature falls within the temperature threshold range again. In some embodiments, the temperature regulator 210 has a function of regulating the operating temperature thereof, for example, a control panel (not shown) may be provided on the incubator 200, and the control panel is connected to the temperature regulator 210 and is used for regulating the operating temperature of the temperature regulator 210. If the temperature of the culture fluid is detected to be higher than 38 ℃, the operating temperature currently set by the temperature regulator 210 can be appropriately adjusted downward so that the internal temperature of the incubator 200 drops and falls within the temperature threshold again. If the temperature of the culture solution is detected to be lower than 36 ℃, the operating temperature currently set by the temperature regulator 210 can be appropriately adjusted to be higher, so that the internal temperature of the incubator 200 rises and falls within the temperature threshold again.

Fig. 10 shows a flow diagram of a temperature adjustment method 3000 for an in vitro experimental system 1000 according to another embodiment of the present disclosure. The temperature adjustment method 3000 may be applied to an in vitro experiment system 1000. The method 3000 includes:

step 3001, setting initial operating temperatures of the air conditioner 500 and the thermostat 210, respectively;

step 3002, detecting the temperature of the culture solution contained in at least one culture dish 110 by using the temperature detector 300;

step 3003, determining whether the detected temperature of the culture medium exceeds an upper threshold, if so, executing step 3004, otherwise, executing step 3006;

step 3004, generating an alarm signal and executing step 3005;

step 3005, setting a target operating temperature of the thermostat 210 based on the difference between the current operating temperature and the temperature of the culture fluid and the desired temperature value, and executing step 3002;

step 3006, determining whether the detected temperature of the culture medium exceeds a lower threshold, if so, executing step 3004; if the determination result is that the detected temperature of the culture solution does not exceed the lower threshold, then step 3007 is executed;

step 3007 continues with repeating step 3002 without changing the temperature setting of thermostat 210.

Prior to step 3001, 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 temperature detector 300 is inserted into the culture solution. In addition, the temperature threshold range may be set to 36 ℃ to 38 ℃ in advance, that is, the alarm upper limit threshold of the alarm of the temperature detector 300 may be set to 38 ℃ and the alarm lower limit threshold may be set to 36 ℃. In step 3001, an initial operating temperature of the incubator 200 may be set to 30 ℃, a carbon dioxide concentration may be set to 5%, and an initial operating temperature of the air conditioner 500 may be set to 24 ℃. After the preparation is completed, the electric field generator 410 is turned on, and the temperature detector 300 continuously detects the temperature of the culture solution.

In step 3003, the upper threshold is 38 ℃.

In step 3004, the alarm signal is sent by an alarm in the temperature detector 300 when the temperature of the cell culture fluid detected by the temperature detector 300 is greater than 38 ℃.

In step 3005, the operating temperature of incubator 200 is adjusted according to the following equation:

Ts＝Tc-Tl+37℃

where Ts is a target operating temperature to be set by the incubator 200, Tc is a current operating temperature currently set by the incubator 200, and Tl is a culture solution temperature. As an example, if the current operating temperature of the incubator 200 is 32 ℃, it is detected during the detection that the culture solution temperature reaches 39 ℃, the alarm gives an alarm signal, and the target operating temperature calculated by the above equation is 30 ℃. Thus, the operating temperature of the thermostat 210 will be adjusted from 32 ℃ to 30 ℃. After the operation temperature of the temperature regulator 210 is adjusted to be low, the temperature of the culture solution will decrease.

In step 3006, the lower threshold is 36 ℃.

When the temperature of the culture solution detected by the temperature detector 300 is less than or equal to 38 ℃, it is determined whether the temperature is less than 36 ℃. If not less than 36 ℃, the temperature setting of thermostat 210 is not changed, i.e., the current operating temperature of thermostat 210 is maintained. Otherwise, the alarm sends out an alarm signal to prompt that the operating temperature of the incubator 200 is adjusted according to the formula.

As an example, if the current operating temperature of the incubator 200 is 32 ℃, and only 35 ℃ of the culture fluid temperature is detected during the detection, the alarm gives an alarm signal, and the target operating temperature calculated using the above equation is 34 ℃. Thus, the operating temperature of the thermostat 210 will be adjusted from 32 ℃ to 34 ℃. After the operation temperature of the temperature regulator 210 is increased, the temperature of the culture solution is increased.

After

steps

3005 and 3007, the above temperature sensing and control of the operating temperature of incubator 200 may be repeated until the temperature of the culture medium tends to stabilize before the alarm is triggered to signal an alarm.

In some embodiments, the above-mentioned adjustment of the operating temperature of the temperature regulator 210 may be automatically performed by a controller, and in one example, the controller may be electrically connected to the temperature detector 300 and the temperature regulator 210, respectively, the temperature detector 300 generates a detection signal after detecting the temperature of the culture solution and transmits the detection signal to the controller, and the controller controls the temperature regulator 210 to adjust the internal temperature of the incubator 200 according to temperature information included in the detection signal.

In other embodiments, the adjustment of the internal temperature of the incubator 200 described above may be performed manually. In one example, the operating temperature of thermostat 210 may be set manually by an associated laboratory operator upon receiving an alarm signal from an alarm.

The in-vitro experiment system 1000 of the invention utilizes the temperature detector 300 to detect the temperature in the culture dish 110, detects that the temperature of the culture solution exceeds the temperature threshold range, adjusts the internal temperature of the culture box 200 by adjusting the temperature adjuster 210 of the culture box 200 to enable the temperature of the culture solution to fall into the temperature threshold range, does not need to adjust alternating voltage, avoids the influence of the fluctuation of the field intensity on the experiment, and is beneficial to the repeatability of the experiment in-vitro research, the quantitative statistics of the experiment result and the like.

The present application is only a preferred embodiment of the present application and should not be limited to the above embodiments, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. An in vitro assay system, comprising:

an incubator comprising a temperature regulator configured to regulate an internal temperature of the incubator;

culture apparatus, set up inside the incubator, include:

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 fluid contained in the culture dish; and

a temperature detector for detecting a temperature of the culture solution contained in the at least one culture dish,

wherein the temperature adjuster adjusts the internal temperature of the incubator so that the culture medium temperature falls within a temperature threshold range when the temperature detector detects that the culture medium temperature exceeds the temperature threshold range.

2. The in vitro assay system of claim 1, wherein the temperature detector comprises:

the tail end of each temperature measuring line is provided with a thermocouple probe which extends into the culture solution in the corresponding culture dish; and

and the test instrument is used for converting the at least one detection signal into a corresponding temperature value.

3. The in vitro assay system of claim 2, wherein the culture device further comprises: and the circuit plugboard 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, wherein the circuit board is provided with a plurality of insertion holes for inserting the at least one set of electrode plates, at least one pin hole for passing a temperature measuring wire is further provided in a region surrounded by the plurality of insertion holes, the at least one culture dish is provided with a corresponding opening at a top end of the at least one culture dish, and the at least one set of electrode plates is arranged on the circuit board through the plurality of insertion holes 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 assay system of claim 4, wherein each set of electrode plates comprises at least two pairs of electrode plates.

6. The in vitro assay system of claim 4, wherein each electrode plate comprises a flexible circuit board, an insulating substrate and a ceramic electrode disposed on opposite sides of the flexible circuit board.

7. The in vitro assay system of claim 6, wherein the flexible circuit board and the insulating substrate are substantially the same shape, and are both disposed substantially in a "T" shape.

8. The in vitro assay system of claim 7, wherein the flexible circuit board has a width at an end thereof proximal to the circuit card that is less than a width at an end thereof distal to the circuit card.

9. The in vitro assay system of claim 6, wherein the ceramic electrode is disposed on an end of the flexible circuit board remote from the circuit board.

10. The in vitro assay system of claim 6, wherein each electrode plate further comprises at least one temperature sensor disposed on an end of the flexible circuit board remote from the circuit board, the temperature sensor being disposed on the same side of the flexible circuit board as the ceramic electrode.

11. The in vitro experimental system of claim 10, wherein each electrode plate further comprises a plurality of conductive terminals disposed at an end of the flexible circuit board adjacent to the circuit board, one end of each conductive terminal is electrically connected to the flexible circuit board, and the other end of each conductive terminal is electrically connected to the circuit board above the culture dish.

12. The in vitro assay system of claim 11, wherein the other end of each conductive terminal is electrically connected to a corresponding jack on the circuit board.

13. The in vitro assay system of claim 3, wherein the culture device further comprises: and the mounting plate assembly is used for mounting and fixing the at least one culture dish and the circuit insertion plate.

14. The in vitro assay system of claim 13, wherein the mounting plate assembly comprises a bottom plate, a plurality of support posts secured to the bottom plate, and a top plate removably secured to the plurality of support posts.

15. The in vitro assay system of claim 14, wherein the base plate is of generally plate-like configuration for supporting at least one culture dish.

16. The in vitro assay system of claim 15, wherein the bottom plate supports one side surface of the culture dish and is provided with at least one positioning member that restricts horizontal movement of the culture dish.

17. The in vitro assay system of claim 14, wherein the top of each support column is provided with a first securing member.

18. The in vitro assay system of claim 17, wherein the top plate is provided with a plurality of second fixing members corresponding to the first fixing members of the plurality of support columns.

19. The in vitro assay system of claim 18, wherein the top plate is removably fixedly coupled to the first securing member of the corresponding support column via a plurality of second securing members thereof.

20. The in vitro assay system of claim 14, wherein the top plate is disposed above the at least one culture dish for supporting a circuit card.

21. The in vitro assay system of claim 20, wherein the top plate defines at least one retaining opening for receiving a respective top end of at least one culture dish.

22. The in vitro experiment system according to claim 3, wherein a flat cable connecting terminal is arranged at one end of the circuit board and used for connecting a flat cable.

23. The in vitro assay system of any one of claims 1 to 22, further comprising: and the electric field generator is used for generating at least one alternating electric field signal and transmitting the alternating electric field signal to at least one group of electrode plates.

24. The in vitro assay system of claim 23, further comprising an adapter electrically connected to the electric field generator for providing the at least one alternating electric field signal to at least one set of electrode plates via the adapter.

25. The in vitro assay system of claim 23, further comprising:

a controller configured to automatically control the temperature regulator to adjust the internal temperature of the incubator such that the culture fluid temperature falls within a temperature threshold range in response to detecting that the culture fluid temperature is outside the temperature threshold range.

26. The in vitro assay system of claim 25, wherein the temperature threshold range is 36 ℃ to 38 ℃.

27. A method of temperature regulation of an in vitro test system according to any one of claims 1 to 26, the method comprising:

periodically detecting the temperature of the culture solution contained in the at least one culture dish by using the temperature detector; and

in response to detecting that the culture fluid temperature is outside a temperature threshold range, controlling the temperature regulator to regulate the internal temperature of the incubator such that the culture fluid temperature falls within the temperature threshold range, the temperature threshold range being 36-38 ℃.

28. The temperature adjustment method of claim 27, wherein the temperature threshold range comprises a desired temperature value at which the culture solution is desired to be maintained, and wherein controlling the temperature regulator to regulate the internal temperature of the incubator further comprises:

acquiring a current operating temperature set for the temperature regulator; and

setting a target operating temperature of the thermostat based on a difference between the current operating temperature and the culture solution temperature and the desired temperature value.

29. The temperature adjustment method according to claim 27 or 28, further comprising:

in response to detecting that the culture fluid temperature does not exceed the temperature threshold range, not changing the temperature setting of the thermostat.