A cell culture device and a cell culture method

Abstract

The present invention discloses a cell culture device and a cell culture method. The inner side of a water bath thermostatic incubator is provided with a groove matching the shape of a flexible culture box body. The water bath thermostatic incubator is a hollow structure. The water bath thermostatic incubator is filled with constant temperature heated water, which is heated by a circulating heating device. The upper edge of the flexible culture box body is fixedly connected to a culture box connecting ring, the lower end of the flexible culture box body is fixedly connected to the edge of a glass culture box bottom plate, the cell culture fluid is located in the flexible culture box body, and the glass culture box cover is detachably fixedly connected to the upper end of the culture box connecting ring. The present invention provides an integrated culture box, so that the cultured cells do not need to be taken out during the culture and observation process, and the culture box automatically acts as a cover glass and a slide glass. During the observation period, the cell activity will not decrease due to the temperature difference, thereby improving the accuracy of the experiment and making the culture observation results accurate.

Description

Technical Field

[0001] The invention relates to biological experimental equipment, in particular to a cell culture device and a cell culture method. Background Art

[0002] Cell culture refers to a method of simulating the in vivo environment (sterility, suitable temperature, pH and certain nutritional conditions, etc.) in vitro to enable it to survive, grow, reproduce and maintain its main structure and function. Cell culture is also called cell cloning technology, and the formal term in biology is cell culture technology. Whether it is for the entire bioengineering technology or one of the biological cloning technologies, cell culture is an indispensable process, and cell culture itself is the large-scale cloning of cells. Cell culture technology can transform a cell into a simple single cell or a multi-cell with very little differentiation through large-scale culture. This is an indispensable part of cloning technology, and cell culture itself is the clone of cells. Cell culture technology is an important and commonly used technology in cell biology research methods. Through cell culture, a large number of cells can be obtained, and cell signal transduction, cell anabolism, cell growth and proliferation, etc. can be studied. In the prior art, cell culture is mostly carried out in culture dishes, and heating and constant temperature need to be carried out in an incubator. After the culture is completed, the cells need to be taken out for observation and then placed on the coverslip and slide of a microscope. The device will come into contact with the air and is easily contaminated. The operation is cumbersome, resulting in inaccurate detection and observation results. In addition, the large temperature difference between the coverslip, the slide and the cultured cells also affects the activity of the cells to a certain extent, thereby reducing the verification accuracy of the culture results. Therefore, there is room for improvement. Summary of the invention

[0003] The object of the present invention is to provide a cell culture device and a cell culture method in order to solve the above problems.

[0004] The present invention achieves the above-mentioned purpose through the following technical solutions:

[0005] The present invention provides a cell culture device, comprising a water bath constant temperature incubator, constant temperature heated water, a culture box connecting ring, a flexible culture box body, a glass culture box bottom plate, a glass culture box cover plate, a cell culture fluid tissue and a circulating heating device, wherein a groove matching the shape of the flexible culture box body is arranged on the inner side of the water bath constant temperature incubator, the water bath constant temperature incubator is a hollow structure, the water bath constant temperature incubator is filled with the constant temperature heated water, the constant temperature heated water is heated by the circulating heating device, the upper end edge of the flexible culture box body is fixedly connected to the culture box connecting ring, the lower end of the flexible culture box body is fixedly connected to the edge of the glass culture box bottom plate, the cell culture fluid tissue is located in the flexible culture box body, the glass culture box cover plate is detachably fixedly connected to the upper end of the culture box connecting ring, and parts of the flexible culture box body and the glass culture box bottom plate are located in the groove of the water bath constant temperature incubator and can be taken out.

[0006] Furthermore, the circulating heating device is composed of a heating circulating water pump, a circulating water heating coil, a coil heating electric heating wire, a circulating water storage tank, and a circulating water return pipe. The water inlet of the water bath constant temperature incubator is connected to the water outlet of the heating circulating water pump, the water outlet of the water bath constant temperature incubator is connected to the water inlet of the circulating water storage tank through the circulating water return pipe, the drain outlet of the circulating water storage tank is connected to the water inlet of the circulating water heating coil, the drain outlet of the circulating water heating coil is connected to the water inlet of the heating circulating water pump, and the coil heating electric heating wire is wound on the surface of the circulating water heating coil.

[0007] Furthermore, the circulation heating device includes a heating temperature sensor, a signal output end of the heating temperature sensor is connected to a signal input end of a heating control system, and a control output end of the heating control system is also connected to the heating circulation water pump and the coil heating heating wire.

[0008] As an improvement, the circulating water heating coil and the coil heating electric heating wire are located in the heating coil insulation shell.

[0009] Preferably, the culture box connecting ring and the glass culture box cover are detachably fixedly connected by adhesive.

[0010] As an improvement, an observation device is also provided, which consists of an observation device base, an observation device light source, an observation device support tube and an electronic observation microscope. The observation device support tube is a cylindrical structure. The observation device base is fixedly arranged at the lower end of the observation device support tube. The middle part of the observation device base is located at the lower end of the observation device support tube. The observation device light source is arranged at the upper end of the observation device support tube. The electronic observation microscope is located above the observation device support tube. The glass culture box bottom plate and the glass culture box cover plate are located at the upper end of the observation device support tube and below the electronic observation microscope.

[0011] Preferably, a light-homogenizing Fresnel lens is provided in the upper section of the support tube of the observation device.

[0012] As an improvement, the water bath constant temperature incubator is provided with a plurality of grooves, and a culture box consisting of the culture box connecting ring, the flexible culture box body, the glass culture box bottom plate and the glass culture box cover plate can be arranged in each groove.

[0013] The present invention provides a cell culture method, comprising the following steps:

[0014] ① When cells need to be cultured, place the cell culture fluid tissue into the flexible culture box body, and bond and seal the glass culture box cover plate and the culture box connecting ring;

[0015] ② Place the culture box consisting of a culture box connecting ring, a flexible culture box body, a glass culture box bottom plate, and a glass culture box cover plate into the groove of a water bath constant temperature incubator;

[0016] ③ The water in the circulating water storage tank is pumped into the water bath constant temperature incubator through the heating circulating water pump to heat the incubator at a constant temperature. During the pumping process, the circulating water heating coil is heated by the coil heating electric heating wire. At the same time, the heating temperature sensor monitors the temperature in the water bath constant temperature incubator. The heating control system controls the heating temperature of the coil heating electric heating wire through the monitoring temperature data of the heating temperature sensor. The water in the water bath constant temperature incubator is then returned to the circulating water storage tank through the circulating water return pipe for storage;

[0017] ④When the cell culture is completed, take out the culture box directly and put it into the observation device for observation;

[0018] ⑤ Since the flexible culture box body of the culture box is flexible, the bottom plate and the cover plate of the glass culture box are both made of glass, the bottom plate of the glass culture box is placed on the support tube of the observation device, and the bottom plate of the glass culture box overlaps with the cover plate of the glass culture box, which is equivalent to the role of the cover glass and the slide glass, and the excess cell culture fluid tissue is located in the flexible culture box body;

[0019] ⑥ The light source of the observation device is powered on to emit light, and the light-averaging Fresnel lens balances the light from the light source. At this time, observation is performed through an electronic observation microscope.

[0020] The beneficial effects of the present invention are:

[0021] The present invention relates to a cell culture device and a cell culture method. Compared with the prior art, the present invention provides an integrated culture box, so there is no need to take out the cultured cells during the culture and observation process, and the culture box automatically acts as a cover glass and a slide. During the observation period, the cell activity will not decrease due to temperature difference, thereby improving the accuracy of the experiment, making the culture observation results accurate, easy to use, and worthy of promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a schematic diagram of the structure of the present invention;

[0023] Figure 2 It is a schematic diagram of the structure of the observation device of the present invention;

[0024] Figure 3 1 is a schematic diagram of the structure of the culture box from a top view;

[0025] Figure 4 It is a schematic diagram of the structure of multiple culture boxes. DETAILED DESCRIPTION

[0026] The present invention will be further described below in conjunction with the accompanying drawings:

[0027] like Figure 1-4 As shown: A cell culture device of the present invention comprises a water bath constant temperature incubator 1, constant temperature heated water 2, a culture box connecting ring 3, a flexible culture box body 4, a glass culture box bottom plate 5, a glass culture box cover plate 6, a cell culture fluid tissue 7 and a circulating heating device, wherein the inner side of the water bath constant temperature incubator 1 is provided with a groove matching the shape of the flexible culture box body 4, the water bath constant temperature incubator 1 is a hollow structure, the water bath constant temperature incubator 1 is filled with the constant temperature heated water 2, and the constant temperature heated water 2 is heated by the circulating heating device, the upper end edge of the flexible culture box body 4 is fixedly connected to the culture box connecting ring 3, the lower end of the flexible culture box body 4 is fixedly connected to the edge of the glass culture box bottom plate 5, the cell culture fluid tissue 7 is located in the flexible culture box body 4, the glass culture box cover plate 6 is detachably fixedly connected to the upper end of the culture box connecting ring 3, and parts of the flexible culture box body 4 and the glass culture box bottom plate 5 are located in the groove of the water bath constant temperature incubator 1 and can be taken out.

[0028] Furthermore, the circulating heating device is composed of a heating circulating water pump 8, a circulating water heating coil 9, a coil heating electric heating wire 10, a circulating water storage tank 12, and a circulating water return pipe 13. The water inlet of the water bath constant temperature incubator 1 is connected to the water outlet of the heating circulating water pump 8, the water outlet of the water bath constant temperature incubator 1 is connected to the water inlet of the circulating water storage tank 12 through the circulating water return pipe 13, the drain outlet of the circulating water storage tank 12 is connected to the water inlet of the circulating water heating coil 9, the drain outlet of the circulating water heating coil 9 is connected to the water inlet of the heating circulating water pump 8, and the coil heating electric heating wire 10 is wound on the surface of the circulating water heating coil 9.

[0029] Furthermore, the circulation heating device includes a heating temperature sensor 14 , a signal output end of the heating temperature sensor 14 is connected to a signal input end of a heating control system 15 , and a control output end of the heating control system 15 is also connected to the heating circulation water pump 8 and the coil heating electric heating wire 10 .

[0030] As an improvement, the circulating water heating coil 9 and the coil heating electric heating wire 10 are located in the heating coil insulation shell 11.

[0031] Preferably, the culture box connecting ring 3 and the glass culture box cover 6 are detachably fixedly connected by adhesive.

[0032] As an improvement, an observation device is also provided, which consists of an observation device base 16, an observation device light source 17, an observation device support tube 18 and an electronic observation microscope 20. The observation device support tube 18 is a cylindrical structure. The observation device base 16 is fixedly arranged at the lower end of the observation device support tube 18. The middle part of the observation device base 16 is located at the lower end of the observation device support tube 18, and the observation device light source 17 is arranged. The electronic observation microscope 20 is located above the observation device support tube 18. The glass culture box bottom plate 5 and the glass culture box cover plate 6 are located at the upper end of the observation device support tube 18 and are located below the electronic observation microscope 20.

[0033] Preferably, a light-homogenizing Fresnel lens 19 is disposed in the upper section of the observation device support tube 18 .

[0034] As an improvement, the water bath constant temperature incubator 1 is provided with a plurality of grooves, and a culture box consisting of the culture box connecting ring 3, the flexible culture box body 4, the glass culture box bottom plate 5, and the glass culture box cover plate 6 can be arranged in each groove.

[0035] The present invention provides a cell culture method, comprising the following steps:

[0036] ① When cells need to be cultured, the cell culture fluid tissue 7 is placed in the flexible culture box body 4, and the glass culture box cover 6 is bonded and sealed with the culture box connecting ring 3;

[0037] ② Place the culture box consisting of the culture box connecting ring 3, the flexible culture box body 4, the glass culture box bottom plate 5, and the glass culture box cover plate 6 into the groove of the water bath constant temperature incubator 1;

[0038] ③ The water in the circulating water storage tank 12 is pumped into the water bath constant temperature incubator 1 through the heating circulating water pump 8 to heat the incubator at a constant temperature. During the pumping process, the circulating water heating coil 9 is heated by the coil heating electric heating wire 10. At the same time, the heating temperature sensor 14 monitors the temperature in the water bath constant temperature incubator 1. The heating control system 15 controls the heating temperature of the coil heating electric heating wire 10 through the monitoring temperature data of the heating temperature sensor 14. The water in the water bath constant temperature incubator 1 is then returned to the circulating water storage tank 12 for storage through the circulating water return pipe 13;

[0039] ④When the cell culture is completed, take out the culture box directly and put it into the observation device for observation;

[0040] ⑤ Since the flexible culture box body 4 of the culture box is flexible, the glass culture box bottom plate 5 and the glass culture box cover plate 6 are both made of glass, the glass culture box bottom plate 5 is placed on the observation device support tube 18, and the glass culture box bottom plate 5 overlaps with the glass culture box cover plate 6, which is equivalent to the role of a cover glass and a slide glass, and the excess cell culture fluid tissue 7 is located in the flexible culture box body 4;

[0041] ⑥ The light source 17 of the observation device is powered on to emit light, and the light-averaging Fresnel lens 19 balances the light from the light source. At this time, observation is performed through the electronic observation microscope 20.

[0042] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A cell culture device, characterized in that: The invention comprises a water bath constant temperature incubator (1), constant temperature heated water (2), a culture box connecting ring (3), a flexible culture box body (4), a glass culture box bottom plate (5), a glass culture box cover plate (6), a cell culture fluid tissue (7) and a circulating heating device, wherein the inner side of the water bath constant temperature incubator (1) is provided with a groove matching the shape of the flexible culture box body (4), the water bath constant temperature incubator (1) is a hollow structure, the water bath constant temperature incubator (1) is filled with the constant temperature heated water (2), the constant temperature heated water (2) is heated by the circulating heating device, the upper end edge of the flexible culture box body (4) is fixedly connected to the culture box connecting ring (3), the lower end of the flexible culture box body (4) is connected to the glass culture box cover plate (6), and the cell culture fluid tissue (7) is connected to the glass culture box body (4). The flexible culture box body (4) is fixedly connected to the edge of the box bottom plate (5), the cell culture fluid tissue (7) is located in the flexible culture box body (4), the glass culture box cover plate (6) is detachably fixedly connected to the upper end of the culture box connecting ring (3), and the flexible culture box body (4) and the glass culture box bottom plate (5) are partially located in the groove of the water bath constant temperature incubator (1) and can be removed; the water bath constant temperature incubator (1) is provided with a plurality of grooves, each of which is provided with a culture box consisting of the culture box connecting ring (3), the flexible culture box body (4), the glass culture box bottom plate (5), and the glass culture box cover plate (6); the culture box connecting ring (3) and the glass culture box cover plate (6) are detachably fixedly connected by adhesive.

2. The cell culture device according to claim 1, characterized in that: The circulating heating device is composed of a heating circulating water pump (8), a circulating water heating coil (9), a coil heating electric heating wire (10), a circulating water storage tank (12), and a circulating water return pipe (13). The water inlet of the water bath constant temperature incubator (1) is connected to the water outlet of the heating circulating water pump (8), the water outlet of the water bath constant temperature incubator (1) is connected to the water inlet of the circulating water storage tank (12) through the circulating water return pipe (13), the water outlet of the circulating water storage tank (12) is connected to the water inlet of the circulating water heating coil (9), the water outlet of the circulating water heating coil (9) is connected to the water inlet of the heating circulating water pump (8), and the coil heating electric heating wire (10) is wound on the surface of the circulating water heating coil (9).

3. The cell culture device according to claim 2, characterized in that: The circulating heating device comprises a heating temperature sensor (14), a signal output end of the heating temperature sensor (14) being connected to a signal input end of a heating control system (15), and a control output end of the heating control system (15) being further connected to the heating circulating water pump (8) and the coil heating electric heating wire (10).

4. The cell culture device according to claim 2, characterized in that: The circulating water heating coil (9) and the coil heating electric heating wire (10) are located in a heating coil insulation shell (11).

5. The cell culture device according to claim 1, characterized in that: An observation device is also provided, the observation device comprising an observation device base (16), an observation device light source (17), an observation device support tube (18) and an electronic observation microscope (20); the observation device support tube (18) is a cylindrical structure; the observation device base (16) is fixedly arranged at the lower end of the observation device support tube (18); the middle part of the observation device base (16) is located at the lower end of the observation device support tube (18); the observation device light source (17) is arranged; the electronic observation microscope (20) is located above the observation device support tube (18); the glass culture box bottom plate (5) and the glass culture box cover plate (6) are located at the upper end of the observation device support tube (18) and below the electronic observation microscope (20).

6. The cell culture device according to claim 5, characterized in that: The upper section of the observation device support tube (18) is provided with a light-homogenizing Fresnel lens (19).

7. A cell culture method, characterized in that: The following steps are involved: ① When cells need to be cultured, the cell culture fluid tissue (7) is placed in the flexible culture box body (4), and the glass culture box cover (6) is bonded and sealed to the culture box connecting ring (3); ② Place the culture box consisting of the culture box connecting ring (3), the flexible culture box body (4), the glass culture box bottom plate (5), and the glass culture box cover plate (6) into the groove of the water bath constant temperature incubator (1); ③ The water in the circulating water storage tank (12) is pumped into the water bath constant temperature incubator (1) by means of a heating circulating water pump (8) to heat the incubator at a constant temperature. During the pumping process, the circulating water heating coil (9) is heated by means of the coil heating electric heating wire (10). At the same time, the heating temperature sensor (14) monitors the temperature in the water bath constant temperature incubator (1). The heating control system (15) controls the heating temperature of the coil heating electric heating wire (10) by means of the temperature data monitored by the heating temperature sensor (14). The water in the water bath constant temperature incubator (1) is then returned to the circulating water storage tank (12) for storage via the circulating water return pipe (13); ④When the cell culture is completed, take out the culture box directly and put it into the observation device for observation; ⑤ Since the flexible culture box body (4) of the culture box is flexible, the glass culture box bottom plate (5) and the glass culture box cover plate (6) are both made of glass, the glass culture box bottom plate (5) is placed on the observation device support tube (18), and the glass culture box bottom plate (5) and the glass culture box cover plate (6) overlap, which is equivalent to the function of a cover glass and a slide glass, and the excess cell culture fluid tissue (7) is located in the flexible culture box body (4); ⑥ The light source (17) of the observation device is powered on to emit light, and the light-averaging Fresnel lens (19) balances the light from the light source. At this time, observation is performed through an electronic observation microscope (20).

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