CN216890992U - In-situ observation cell culture device - Google Patents

Abstract

The utility model provides an in-situ observation cell culture device, which comprises a culture box and a control box communicated with the culture box, wherein the culture box comprises a bottom plate, a top plate, and an inner layer plate, a middle layer plate and an outer layer plate which are sequentially arranged between the bottom plate and the top plate in a surrounding manner from inside to outside; a culture chamber is arranged in the inner-layer plate; a heating plate is arranged between the inner layer plate and the middle layer plate; the inner layer plate, the middle layer plate and the outer layer plate are all provided with through holes; a plurality of sensors connected with the control box are arranged in the culture box; the sensor is used for transmitting the acquired sensing signals to the control box; the control box is used for providing control signals and gas flow for the culture box according to the sensing signals; the control box transmits the gas flow to enter the culture chamber after sequentially passing through the outer plate, the middle plate, the heating plate and the inner plate. The utility model ensures that the thermal uniformity in the culture chamber is better, and can effectively improve the temperature and the gas environment in the cell culture chamber.

Description

In-situ observation cell culture device

Technical Field

The utility model belongs to the technical field of cell culture, and particularly relates to an in-situ observation cell culture device.

Background

The living cells are basic structural units and functional units of life bodies, so that the living cells have important significance for real-time observation of single living cells. In order to observe activities, migration, proliferation and other behaviors of cells in real time for a long time, a small movable cell culture device matched with a microscope stage mechanism is required to realize time-delay sequence acquisition of image data.

However, in the current in-situ observation cell culture device, gas mixing is completed outside the culture box, and then the gas is blown into the culture box by using a fan, which ensures uniformity and concentration of the gas in a mixing area, but the gas flow path is subjected to more links, so that the cost of heating the gas and sealing the gas is increased, and the probability of gas leakage is also increased; the difference between the concentration of the gas after it reaches the terminal region of the incubator and the predetermined concentration of the mixing zone is increased.

In addition, although the in-situ observation of the unicellular culture process can be realized by part of the object stage culture devices, the bottom heat preservation effect is not good, or the internal thermal uniformity is not good, and the problems that the connection of the circuit and the pipeline of the control box and the culture box is not convenient and fast exist.

SUMMERY OF THE UTILITY MODEL

The utility model aims to overcome the defects in the prior art and provide an in-situ observation cell culture device which can effectively improve the temperature and the gas environment inside a cell culture chamber.

In order to achieve the purpose, the utility model is realized by adopting the following technical scheme:

the utility model provides an in-situ observation cell culture device, which comprises a culture box and a control box communicated with the culture box;

the culture box comprises a bottom plate, a top plate, and an inner layer plate, a middle layer plate and an outer layer plate which are sequentially arranged between the bottom plate and the top plate in an enclosing manner from inside to outside; a culture chamber is arranged in the inner-layer plate; a heating plate is arranged between the inner layer plate and the middle layer plate; the inner layer plate, the middle layer plate and the outer layer plate are all provided with through holes;

a plurality of sensors connected with the control box are arranged in the culture box; the sensor is used for transmitting the acquired sensing signals to the control box;

the control box is used for providing control signals and gas flow for the culture box according to the sensing signals; the control box transmits the gas flow to enter the culture chamber after sequentially passing through the outer plate, the middle plate, the heating plate and the inner plate.

The effect achieved by the above arrangement is as follows: the culture box of the in-situ observation cell culture device provided by the utility model is provided with three layers of walls, namely an inner layer plate, a middle layer plate and an outer layer plate, which are respectively matched with a top plate and a bottom plate to form relatively closed spaces, gas injection and mixing are completed in the culture box, and gas enters the inner layer from the middle layer, so that the process is smoother, the thermal uniformity in the culture chamber is better, the temperature environment in the cell culture chamber can be effectively improved, the temperature control level in the cell culture chamber is improved, and the culture and test effects are effectively ensured.

Furthermore, a master access hole is formed in the outer plate, and an air pipe and a lead which are communicated with the control box penetrate through the master access hole; the middle layer plate is provided with an air inlet hole for communicating an air pipe and a power line inlet hole for penetrating a lead;

the inner-layer plates are all heat dissipation plates; the inner plate is provided with at least one group of air hole arrays.

The effect achieved by the above arrangement is: the hot plate is located between the inner plate and the middle plate of cultivation box, and whole inner plate is as the heating panel of hot plate for heat-conducting area increases, and then makes hot homogeneity better. Through-hole array on the heating panel for it switches on gas with higher speed, and gaseous inlet port is located a wallboard of plywood in the culture box, after the trachea from getting into the inlet port through total inlet hole, gaseous diffusion is carried out along the clearance of enclosing between culture box inner plate and the plywood to the gas pocket array on two heating panels through the inner plate gets into the culture chamber, and then makes the cell sample who cultivates in the chamber obtain good cultivation and regulation and control environment.

Further, the air hole array comprises a plurality of air holes with the diameter of a single hole being less than 2 mm.

The effect achieved by the above arrangement is: the gas hole diameter is little and many, has both been favorable to the inside and outside heat transfer of inner plating, also is favorable to making gaseous fully to diffuse between inner plating and outer plywood, and in addition, gaseous before getting into the interior chamber of cultivateing the box, will pass through the round clearance between middle plating and the inner plating, and the inner plating is the heating panel of hot plate, and it can preheat gas, utilizes the convection heat transfer effect that the gas flow process arouses to make heat transfer efficiency and hot homogeneity improve.

Furthermore, four fastening knobs are arranged on the top plate and used for hermetically connecting the middle-layer plate and the top plate.

The effect achieved by the above arrangement is: the fastening knob is used to fasten the top plate to the underlying middle plate so that the interior chamber is sealed from air.

Further, a bottom observation hole is formed in the bottom plate; the bottom observation hole is detachably connected with a reducing hose through a flange; the lower end of the reducing hose is detachably connected with a straight hose; the bottom observation hole is connected with an objective lens of the inverted microscope through a reducing hose; the diameter of the straight hose is the same as that of the objective lens matched with the straight hose, and the straight hose is in sealing fit with the cylindrical surface of the objective lens.

The effect achieved by the above arrangement is: the peripheral area of the bottom observation hole of the culture box is connected with the flange surface in a sealing mode through bolts, the lower end of the flange structure is connected with the variable-diameter hose, the variable-diameter hose is made of elastic materials, and the diameter of the straight hose is the same as that of the objective lens matched with the straight hose, so that the straight hose tightly wraps the objective lens and air leakage is prevented. Through the bottom observation hole and the reducing hose, the device can realize in-situ microscopic observation and cell real-time image acquisition in the single cell culture process, research the long-time dynamic behavior of cells, and can be matched with a controllable mobile station for use, so that the simultaneous culture, observation and image acquisition of a plurality of groups of cells are realized, and the system robustness is improved.

Further, the top plate is a transparent plate.

The effect achieved by the above arrangement is: the transparent plate facilitates observation and allows the glazing of an inverted microscope to shine through the transparent plate onto the cell sample inside the culture box.

Furthermore, a sample placing area for placing the cell culture plate and the culture disc and an embedded mechanism for installing the cell culture plate and the culture disc are arranged in the culture chamber.

The effect achieved by the above arrangement is: after the top plate is opened, the cell culture plate can be easily placed into the cell culture plate through the embedding mechanism, and the cell culture plate is favorably fixed and observed.

Further, the sensors comprise a temperature sensor arranged in the culture chamber and a gas concentration sensor arranged between the inner layer plate and the middle layer plate;

the control box comprises an upper cover, a middle cover and a bottom cover which are connected from top to bottom;

the upper cover is of a box-shaped structure with an opening at the lower end;

the middle cover is of a box-shaped structure with an opening at the upper end, and the rear panel of the middle cover extends downwards to be connected with the bottom cover;

the bottom cover is of a U-shaped structure with an open front and back, and the front end of the bottom cover is detachably connected with a bottom cover front panel; a direct current power supply pack, a deconcentrator, a gas filter and a gas flow control valve arranged at the input end of the gas filter are arranged in the bottom cover;

the interior of the middle cover is provided with: the gas concentration sensing device comprises a gas concentration sensing signal control plate, a gas concentration PID controller, a temperature PID controller, a gas relay and a hot plate relay;

the gas concentration sensing signal control board is connected with the gas concentration sensor through a wire so as to obtain the gas concentration in the culture chamber, and the output end of the gas concentration sensing signal control board is connected with the input end of the gas concentration PID controller; the output end of the gas concentration PID controller is connected with a gas flow control valve through the gas relay and is used for controlling the gas inlet flow;

the temperature PID controller is connected with the temperature sensor through a wire to acquire the temperature in the culture chamber; the temperature PID controller is connected with the heating plate through a heating plate relay and is used for controlling the on-off of the current of the heating plate;

the output end of the direct current power supply pack is divided into a plurality of paths through a deconcentrator to supply power for the gas concentration sensing signal control plate, the gas concentration PID controller, the temperature PID controller and the heating plate; the gas filter is used for filtering gas, one end of the gas filter is connected with the gas tank through a gas flow control valve, and the other end of the gas filter is communicated with the master inlet hole of the culture box through a gas pipe.

The effect achieved by the above arrangement is: through the control box, can real time control cultivate temperature and gas flow in the chamber, convenient to use controls accurately. The wiring of control box and cultivation box inside all is accomplished at each interlayer of box inside, and wiring and pipeline are more regular, and cultivates the pipeline integration between box and the control box and be a branch of, and it is more convenient to remove the cultivation box.

Furthermore, a fan for accelerating the circulation of air is also arranged in the middle layer plate of the culture box;

a fan operation control panel connected with the fan through a relay is also arranged in the middle cover; the fan operation control panel is connected with the direct current power supply group and used for controlling the fan to operate.

The effect achieved by the above arrangement is: the fan is favorable to driving the circulation of the gas inside the culture box, so that the cell sample in the culture chamber can be well cultured and the environment can be well regulated and controlled.

Furthermore, a side panel of the upper cover and a side panel of the middle cover are provided with a concave-convex block nesting structure which is matched with each other and is used for connecting the upper cover and the middle cover; the front panel of the upper cover is matched with the front panel of the middle cover to form a gap for installing the gas concentration PID controller and the temperature PID controller;

and the gas concentration PID controller and the temperature PID controller are both provided with display screens for displaying information in the culture chamber and inputting control instructions.

The effect achieved by the above arrangement is as follows: the concave-convex block nesting structure is favorable for connecting and installing the upper cover and the middle cover, and the matched notch is favorable for installing the gas concentration PID controller and the temperature PID controller and exposing the display screen.

Furthermore, through holes are formed in the rear panel or the two side panels of the middle cover and used for the inlet and outlet of the conducting wires and the gas pipeline; and the bottom of the bottom cover or the side panel is provided with a threaded hole for mounting parts.

The effect achieved by the above arrangement is: the wiring of control box inside all accomplishes at each interlayer and the screw hole of box inside, and wiring and pipeline are more regular, and cultivates the pipeline integration between box and the control box and be a branch of, and it is more convenient to remove the cultivation box.

Furthermore, reinforcing columns are arranged on the inner sides of four corners of the bottom cover, and the columns are fastened with two adjacent vertical panels through bolts;

the upper and lower positions of the bolt between the reinforcing column body and the two connected panels are staggered;

the effect achieved by the above arrangement is as follows: strengthen cylinder and bolt position and can make two panels and overall structure's that become the right angle intensity better, should not warp the damage.

Compared with the prior art, the utility model has the following beneficial effects:

1. the in-situ observation cell culture device provided by the utility model has the advantages that the culture box is provided with the three walls of the inner layer plate, the middle layer plate and the outer layer plate, the injection and the mixing of gas are completed in the culture box, the gas enters the inner layer plate from the middle layer plate, the process is more gentle, and the thermal uniformity in the culture chamber is better;

2. before entering the inner chamber of the culture box, the gas passes through a circle of gap between the middle-layer plate and the inner-layer plate, the inner-layer plate is a heat dissipation plate of the heating plate and can preheat the gas, and the heat transfer efficiency and the heat uniformity are improved by utilizing the convection heat transfer effect caused by the gas flowing process.

3. The wiring of control box and cultivation box inside all is accomplished at each interlayer of box inside, and wiring and pipeline are more regular, and cultivates the pipeline integration between box and the control box and be a branch of, and it is more convenient to remove the cultivation box.

4. The device can realize in-situ microscopic observation in the process of single cell culture and simultaneously acquire real-time images of cells, research the long-time dynamic behavior of the cells, and simultaneously can be matched with a controllable mobile station for use, so that simultaneous culture, observation and image acquisition of a plurality of groups of cells are realized, and the robustness of the system is improved.

Drawings

FIG. 1 is a schematic view showing a structure of a culture cassette according to a second embodiment of the present invention;

FIG. 2 is a schematic view showing a bottom tubular connection structure of a culture cassette according to a second embodiment of the present invention.

FIG. 3 is a schematic view showing the structure of the inner chamber, inner, middle and outer partitions of the culture box according to the second embodiment of the present invention;

FIG. 4 is a schematic view showing the positional relationship between the air guide hole of the wire and the inner, middle and outer layer structures of the culture box according to the second embodiment of the present invention;

fig. 5 is an exploded view of the control box according to the second embodiment of the present invention;

fig. 6 is a schematic view of an installation relationship of internal component structures of a control box according to a second embodiment of the present invention.

In the figure: 1. an upper cover; 2. a middle cover; 3. a bottom cover; 21. a gas concentration PID controller; 22. a temperature PID controller; 23. a gas concentration sensing signal control panel; 24. a fan operation control panel; 25. the upper cover and the middle cover are connected through the wire connecting through hole; 26. the wire behind the middle cover is connected with the through hole; 31. a DC power pack; 32. a wire divider; 33. a bottom cover front panel; 34. an expanding hole; 41. an inner layer board; 42. a middle layer plate; 420. a power line inlet; 421. an air intake; 43. an outer plate; 44. a base plate; 45. a flange; 46. a variable diameter hose; 47. a straight hose; 48. an access hole; 5. a sample placement area; 51. and (4) embedding the mechanism.

Detailed Description

The utility model is further described below with reference to the accompanying drawings. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.

In the description of the present embodiment, it should be noted that, as the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. appear, the indicated orientation or positional relationship thereof is based on the orientation or positional relationship shown in the drawings, and is only for convenience of describing the present embodiment and simplifying the description, but does not indicate or imply that the referred device or element must have a specific orientation, be configured and operated in a specific orientation, and thus, cannot be construed as limiting the present embodiment.

The first embodiment is as follows:

the embodiment provides an in-situ observation cell culture device, which comprises a culture box and a control box communicated with the culture box, as shown in FIGS. 1 to 4; the culture box comprises a bottom plate 44, a top plate, and an inner layer plate 41, a middle layer plate 42 and an outer layer plate 43 which are sequentially arranged between the bottom plate 44 and the top plate in a surrounding manner from inside to outside; a culture chamber is arranged in the inner layer plate 41; a heating plate is arranged between the inner layer plate 41 and the middle layer plate 42; the inner layer plate 41, the middle layer plate 42 and the outer layer plate 43 are all provided with through holes; a plurality of sensors connected with the control box are arranged in the culture box; the sensor is used for transmitting the acquired sensing signal to the control box; the control box is used for providing control signals and gas flow for the culture box according to the sensing signals; the control box transmits the gas flow to enter the culture chamber after passing through the outer plate 43, the middle plate 42, the heating plate and the inner plate 41 in sequence.

The implementation principle is as follows: the culture box of the in-situ observation cell culture device provided by the embodiment has an inner layer plate 41, an intermediate layer plate 42, three layers of walls of an outer layer plate 43, cooperate with a top plate and a bottom plate 44 respectively to form a relatively closed space, the injection and the mixing of gas are completed in the culture box, the intermediate layer enters into the inner layer, the process is more gentle, the heat uniformity inside the culture chamber is better, the temperature environment inside the cell culture chamber can be effectively improved, the temperature control level in the cell culture chamber is improved, and the culture and test effects are effectively guaranteed.

When the culture box is used, the gas flow is firstly conveyed to the culture box through the control box, the gas flow firstly passes through the through holes of the outer layer plate 43 and is conveyed into the middle layer plate 42 through the gas pipe, a relatively closed space is formed between the inner layer plate 42 and the top plate, the bottom plate 44 and the inner layer plate 41, the gas is heated through the heating plate, and then the heated gas enters the culture cavity through the through holes on the inner layer plate 41.

Example two:

the embodiment provides an in-situ observation cell culture device which comprises two parts, namely a culture box (shown in figures 1-4) and a control box (shown in figures 5-6).

As shown in FIGS. 1 to 4, the culture box comprises a bottom plate 44, a top plate, and an inner plate 41, a middle plate 42 and an outer plate 43 which are sequentially arranged between the bottom plate 44 and the top plate from inside to outside; a culture chamber is arranged in the inner layer plate 41; a heating plate is arranged between the inner layer plate 41 and the middle layer plate 42; the inner plate 41, the middle plate 42 and the outer plate 43 are all provided with through holes.

A plurality of sensors connected with the control box are arranged in the culture box; the sensor is used for transmitting the acquired sensing signal to the control box. Specifically, the sensors include a temperature sensor disposed in the culture chamber and a gas concentration sensor disposed between the inner plate 41 and the middle plate 42, which are connected to the control box through wires, respectively. The remainder of the sensor can span the area between the middle plate 42 to the outer plate 43. The sensor has no fixed installation position, and can be changed into various types as long as the sensor meets the requirements.

The control box is used for providing control signals and gas flow for the culture box according to the sensing signals; the control box transmits the gas flow to enter the culture chamber after passing through the outer plate 43, the middle plate 42, the heating plate and the inner plate 41 in sequence.

The culture box is provided with three walls of an inner layer plate 41, a middle layer plate 42 and an outer layer plate 43 which are respectively matched with a top plate and a bottom plate 44 to form a relatively closed space, gas injection and mixing are completed in the culture box, the gas enters the inner layer from the middle layer, the process is smoother, and the thermal uniformity inside the culture chamber is better.

Preferably, the outer plate 43 is provided with a master access hole 48, and a gas pipe and a lead wire for communicating with the control box are arranged in the master access hole 48 in a penetrating manner. The middle layer plate 42 is provided with an air inlet 421 for communicating an air pipe and a power line inlet 420 for leading a lead; the inner plates 41 are all heat dissipation plates; at least one air hole array is arranged on the inner plate 41.

Specifically, the inner plate 41 is surrounded by four heat dissipation plates, and the whole inner plate 41 serves as a heat dissipation plate of the heating plate, so that the area of heat conduction is increased, and the heat uniformity is better (fig. 3). The inner plate 41 has through hole arrays on the heat dissipation plates of two of its short sides for communicating gas with higher speed, and the gas inlet 421 is located on a wall plate of the middle plate 42 in the culture box (fig. 4), after the gas hose enters the gas inlet through the total inlet hole 48, the gas diffuses along the gap enclosed between the inner plate 41 and the middle plate 42 in the culture box, and enters the culture chamber through the hole arrays on the two heat dissipation plates of the inner plate 41, thereby making the cell sample in the culture chamber obtain good culture and regulation environment. The total inlet holes 48 have a single diameter greater than 1cm and the array of air holes is an array of through holes having a diameter less than 2 mm.

The heating plate is located between the inner plate 41 and the middle plate 42 of the culture box, and the whole inner plate 41 is used as a heat dissipation plate of the heating plate, so that the area of heat conduction is increased, and further the heat uniformity is better. The through hole array on the heating panel is used for conducting gas in an accelerating mode, the gas inlet hole is located in one wall plate of the middle layer plate 42 in the culture box, after the gas pipe enters the gas inlet hole through the master inlet hole 48, the gas is diffused along a gap formed by surrounding of the inner layer plate 41 and the middle layer plate 42 of the culture box, and enters the culture chamber through the gas hole array on the two heating panels of the inner layer plate 41, so that cell samples in the culture chamber can be well cultured and the environment can be well regulated and controlled.

The diameter of the air holes is small and many, which is not only beneficial to the heat transfer inside and outside the inner plate 41, but also beneficial to the gas to be fully diffused between the inner plate 41 and the outer plate 43, in addition, the gas passes through a circle of gap between the middle plate 42 and the inner plate 41 before entering the inner chamber of the culture box, the inner plate 41 is a heat dissipation plate of the heating plate, the gas can be preheated, and the heat transfer efficiency and the heat uniformity are improved by utilizing the convection heat transfer effect caused by the gas flowing process.

Preferably, four fastening knobs are provided on the top plate for hermetically connecting the middle plate 42 with the top plate. The fastening knob is used to fasten the top plate to the underlying middle plate 42 so that the interior chamber is sealed from air.

Preferably, as shown in fig. 2, the bottom plate 44 is provided with a bottom viewing hole; the bottom observation hole is detachably connected with a reducing hose 46 through a flange 45; the lower end of the reducing hose 46 is detachably connected with a straight hose 47; the bottom observation hole is connected with the objective lens of the inverted microscope through a reducing hose 46; the straight hose 47 has the same diameter as the mating objective lens.

In cell culture, the bottom viewing port may be sealed with a removable mechanism, such as a removable sealing plate attached via flange 45.

As shown in figure 2, the peripheral area of the bottom observation hole of the culture box is hermetically connected with the surface of the flange 45 through bolts, the lower end of the flange 45 is connected with the reducing hose 46, the reducing hose 46 is made of elastic material, and the diameter of the straight hose 47 is the same as that of the objective lens matched with the reducing hose, so that the straight hose 47 tightly wraps the objective lens to prevent air leakage. Through the bottom observation hole and the reducing hose 46, the device can realize in-situ microscopic observation and cell real-time image acquisition in the process of single cell culture, research the long-time dynamic behavior of cells, and can be matched with a controllable mobile station for use, so that the simultaneous culture, observation and image acquisition of a plurality of groups of cells are realized, and the system robustness is improved.

Specifically, the top plate is a transparent plate. The transparent plate facilitates observation and allows the glazing of an inverted microscope to shine through the transparent plate onto the cell sample inside the culture box.

Preferably, a sample placement area 5 for placing a cell culture plate and an embedded mechanism 51 for mounting a cell culture plate are provided in the culture chamber.

After the top plate is opened, the cell culture plate can be easily placed through the embedding mechanism 51, which is beneficial to fixing and observing the cell culture plate.

The culture box has three layers of an inner layer plate 41, a middle layer plate 42 and an outer layer plate 43, the heating plate is positioned on the inner layer plate 41 of the culture box, and the sample placing area 5 in fig. 3 is a placing area of a cell culture plate (a plurality of holes are arranged in the culture plate, and cells are implanted in each hole), if a circular culture plate with the diameter of 100mm is adopted, the culture plate can be embedded into a pair of embedding mechanisms 51 with the shape similar to a crescent in the sample placing area 5.

Specifically, as shown in fig. 5-6, the control box comprises an upper cover 1, a middle cover 2 and a bottom cover 3 which are connected from top to bottom;

the upper cover 1 is of a box-shaped structure with an opening at the lower end; the middle cover 2 is a box-shaped structure with an opening at the upper end, and the rear panel extends downwards to be connected with the bottom cover 3; the bottom cover 3 is a U-shaped structure with an open front and back, and the front end of the bottom cover is detachably connected with a bottom cover front panel 33; the bottom cover 3 is internally provided with a direct current power supply group 31, a deconcentrator 32, a gas filter and a gas flow control valve arranged at the input end of the gas filter.

The inside of well lid 2 is provided with: a gas concentration sensing signal control plate 23, a gas concentration PID controller 21, a temperature PID controller 22, a gas relay and a hot plate relay; the gas concentration sensing signal control plate 23 is connected with a gas concentration sensor through a lead to acquire the gas concentration in the culture chamber, and the output end of the gas concentration sensing signal control plate 23 is connected with the input end of the gas concentration PID controller 21; the output end of the gas concentration PID controller 21 is connected with a gas flow control valve through a gas relay and is used for controlling the gas inlet flow; the temperature PID controller 22 is connected with the temperature sensor through a lead to acquire the temperature in the culture chamber; the temperature PID controller 22 is connected with the heating plate through a heating plate relay and is used for controlling the current on-off of the heating plate.

The output end of the direct current power supply group 31 is divided into a plurality of paths through a deconcentrator 32 to supply power for the gas concentration sensing signal control plate 23, the gas concentration PID controller 21, the temperature PID controller 22 and the heating plate; the gas filter is used for filtering air, one end of the gas filter is connected with the gas tank through a gas flow control valve, and the other end of the gas filter is communicated with a total inlet hole 48 of the culture box through a gas pipe.

Through the control box, can real time control cultivate temperature and gas flow in the chamber, convenient to use controls accurately. The wiring of control box and cultivation box inside all accomplishes at each interlayer of box inside, and wiring and pipeline are more regular, and cultivates the pipeline integration between box and the control box and be a branch of, and it is more convenient to remove the cultivation box.

Preferably, a fan for accelerating the circulation of air is further arranged in the middle layer plate 42 of the culture box; a fan operation control panel 24 connected with the fan through a relay is also arranged in the middle cover 2; the fan operation control board 24 is connected to the dc power supply group 31 for controlling the operation of the fan. The fan is favorable to driving the circulation of the gas inside the culture box, so that the cell sample in the culture chamber can be well cultured and the environment can be well regulated and controlled. The micro fan can be arranged between the inner layer and the middle layer or in the inner cavity without fixed position.

Preferably, the side panels of the upper cover 1 and the side panels of the middle cover 2 are provided with a concave-convex block nesting structure matched with each other for connecting the upper cover 1 and the middle cover 2; the front panel of the upper cover 1 is provided with a gap for installing a gas concentration PID controller 21 and a temperature PID controller 22 in a matching way with the front panel of the middle cover 2; the gas concentration PID controller 21 and the temperature PID controller 22 are both provided with display screens for displaying information in the culture chamber and inputting control instructions.

The concave-convex block nesting structure is beneficial to the connection and installation of the upper cover 1 and the middle cover 2, and the matched gap is beneficial to installing the gas concentration PID controller 21 and the temperature PID controller 22 and exposing the display screen.

Specifically, through holes are formed in the rear panel or two side panels of the middle cover 2 and used for leading wires and gas pipelines to enter and exit; the bottom or the side panel of the bottom cover 3 is provided with a threaded hole for mounting parts. The wiring of control box inside all accomplishes at each interlayer and the screw hole of box inside, and wiring and pipeline are more regular, and cultivates the pipeline integration between box and the control box and be a branch of, and it is more convenient to remove the cultivation box. As shown in fig. 5 to 6, includes wire connecting through holes 25 of the upper and middle caps 1 and 2 and wire connecting through holes 26 provided at the rear of the middle cap 2 for connecting wires.

Preferably, as shown in fig. 1, the inner sides of the four corners of the bottom cover 3 are provided with reinforcing columns, and the columns and two adjacent vertical panels are fastened by bolts; the upper and lower positions of the bolt between the reinforcing column body and the two connected panels are staggered; strengthen cylinder and bolt position and can make two panels and overall structure's that become the right angle intensity better, should not warp the damage.

In addition, the control box is also provided with an expansion hole 34 for expanding the functional module.

The implementation process comprises the following steps: the gas line from the gas generator (commercially available, and may be a gas tank) first enters the control box, passes through the gas flow control valve (the valve is controlled by a corresponding gas relay whose frequency is controlled by a PID controller), passes through the filter, exits the control box, and then enters the inlet port 48 of the incubator. After the gas hose enters the gas inlet hole through the total inlet hole 48, gas diffuses along a gap enclosed between the inner plate 41 and the middle plate 42 of the culture box and enters the inner cavity through the hole arrays on the two heat dissipation plates of the inner plate 41, so that cell samples in the inner cavity are well cultured and regulated, the temperature and the gas environment inside the cell culture chamber are effectively improved, and the cell culture quality and the observation effect are improved.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature, and in the description of the utility model, "plurality" means two or more unless explicitly specifically defined otherwise.

In the present invention, unless otherwise specifically stated or limited, the terms "mounted," "connected," "fixed," and the like are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

In the present invention, unless otherwise expressly stated or limited, "above" or "below" a first feature means that the first and second features are in direct contact, or that the first and second features are not in direct contact but are in contact with each other via another feature therebetween. Also, the first feature being "on," "above" and "over" the second feature includes the first feature being directly on and obliquely above the second feature, or merely indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly above and obliquely above the second feature, or simply meaning that the first feature is at a lesser level than the second feature.

In the description herein, reference to the terms "one embodiment," "some embodiments," "an example," "a specific example," or "some examples" or the like means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the utility model. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made in the above embodiments by those of ordinary skill in the art without departing from the principle and spirit of the present invention.

Claims (10)

1. An in-situ observation cell culture device is characterized by comprising a culture box and a control box communicated with the culture box;

the culture box comprises a bottom plate, a top plate, and an inner layer plate, a middle layer plate and an outer layer plate which are sequentially arranged between the bottom plate and the top plate in an enclosing manner from inside to outside; a culture chamber is arranged in the inner-layer plate; a heating plate is arranged between the inner layer plate and the middle layer plate; the inner layer plate, the middle layer plate and the outer layer plate are all provided with through holes;

a plurality of sensors connected with the control box are arranged in the culture box; the sensor is used for transmitting the acquired sensing signals to the control box;

the control box is used for providing control signals and gas flow for the culture box according to the sensing signals; the control box transmits the gas flow to enter the culture chamber after sequentially passing through the outer plate, the middle plate, the heating plate and the inner plate.

2. The in-situ observation cell culture device according to claim 1, wherein a total inlet hole is arranged on the outer plate, and an air pipe and a lead wire for communicating with the control box are arranged in the total inlet hole; the middle layer plate is provided with an air inlet hole for communicating an air pipe and a power line inlet hole for leading through a lead;

the inner-layer plates are all heat dissipation plates; the inner plate is provided with at least one group of air hole arrays.

3. The in situ observation cell culture device of claim 2, wherein the air vent array comprises a plurality of air vents with a single hole diameter of less than 2 mm;

and the top plate is provided with four fastening knobs for hermetically connecting the middle-layer plate and the top plate.

4. The in situ observation cell culture device of claim 1, wherein the bottom plate is provided with a bottom observation hole; the bottom observation hole is detachably connected with a reducing hose through a flange; the lower end of the reducing hose is detachably connected with a straight hose; the bottom observation hole is connected with an objective lens of the inverted microscope through a reducing hose; the diameter of the straight hose is the same as that of the objective lens matched with the straight hose, and the straight hose is in sealing fit with the cylindrical surface of the objective lens;

the top plate is a transparent plate.

5. The in situ cell culture device according to claim 1, wherein a sample placing area for placing the cell culture plate and the culture tray and an embedded mechanism for installing the cell culture plate and the culture tray are arranged in the culture chamber.

6. The in situ cell culture device as recited in claim 2, wherein the sensors comprise a temperature sensor disposed within the culture chamber and a gas concentration sensor disposed between the inner plate and the middle plate;

the control box comprises an upper cover, a middle cover and a bottom cover which are connected from top to bottom;

the upper cover is of a box-shaped structure with an opening at the lower end;

the middle cover is of a box-shaped structure with an opening at the upper end, and the rear panel of the middle cover extends downwards to be connected with the bottom cover;

the bottom cover is of a U-shaped structure with an open front and back, and the front end of the bottom cover is detachably connected with a bottom cover front panel; a direct current power supply pack, a deconcentrator, a gas filter and a gas flow control valve arranged at the input end of the gas filter are arranged in the bottom cover;

the interior of the middle cover is provided with: the gas concentration sensing device comprises a gas concentration sensing signal control plate, a gas concentration PID controller, a temperature PID controller, a gas relay and a hot plate relay;

the gas concentration sensing signal control board is connected with the gas concentration sensor through a wire so as to obtain the gas concentration in the culture chamber, and the output end of the gas concentration sensing signal control board is connected with the input end of the gas concentration PID controller; the output end of the gas concentration PID controller is connected with a gas flow control valve through the gas relay and is used for controlling the gas inlet flow;

the temperature PID controller is connected with the temperature sensor through a wire to acquire the temperature in the culture chamber; the temperature PID controller is connected with the heating plate through a heating plate relay and is used for controlling the on-off of the current of the heating plate;

the output end of the direct current power supply pack is divided into a plurality of paths through a deconcentrator to supply power for the gas concentration sensing signal control plate, the gas concentration PID controller, the temperature PID controller and the heating plate; the gas filter is used for filtering gas, one end of the gas filter is connected with the gas tank through a gas flow control valve, and the other end of the gas filter is communicated with the main inlet hole of the culture box through a gas pipe.

7. The in-situ observation cell culture device according to claim 6, wherein a fan for accelerating the circulation of air is further disposed inside the middle layer plate of the culture box;

a fan operation control panel connected with the fan through a relay is also arranged in the middle cover; the fan operation control panel is connected with the direct current power supply group and used for controlling the fan to operate.

8. The in-situ observation cell culture device of claim 6, wherein the side panels of the upper cover and the side panels of the middle cover are provided with a matched concave-convex block nesting structure for connecting the upper cover and the middle cover; the front panel of the upper cover is matched with the front panel of the middle cover to form a gap for installing the gas concentration PID controller and the temperature PID controller;

and the gas concentration PID controller and the temperature PID controller are both provided with display screens for displaying information in the culture chamber and inputting control instructions.

9. The in-situ observation cell culture device of claim 6, wherein the rear panel or both side panels of the middle cover are provided with through holes for the ingress and egress of wires and gas pipelines; and the bottom of the bottom cover or the side panel is provided with a threaded hole for mounting parts.

10. The in situ observation cell culture device of claim 9, wherein the bottom cover is provided with reinforcing columns at the inner sides of four corners, and the columns are fastened with two adjacent vertical panels by bolts;

the up-down positions of the reinforcing column and the bolt between the two connected panels are staggered.

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