CN102229888B - Feedback-type pneumatic-control pressure stress cell culture device - Google Patents

Abstract

The invention relates to a feedback air-controlled pressure stress cell culture device. The high-pressure gas storage tank of the present invention passes through a switch, a pressure reducing valve, a connecting conduit to a three-way pipe a, and a lower connecting conduit, a connecting conduit, a syringe connected to a micro pressure pump, a connecting conduit to a three-way pipe b, and connecting conduits and a pressure gauge And the upper end of the sterilizing filter, the lower end of the sterilizing filter is connected to the needle; the rubber stopper is on the measuring cylinder glass container, the upper outer reinforcing plate is on the rubber stopper, the lower outer reinforcing plate is under the cylindrical glass container, the needle penetrates the rubber stopper and enters the measuring cylinder Inside the glass container, place the cell culture dish at the bottom of the graduated cylinder glass container. The invention solves the defect that the airtight container is not easy to open and close and cannot withstand a certain amount of compressive stress. The invention cleverly combines a micro pressure pump and a compressive stress cell culture device to provide a compressive stress cell culture device with simple structure, convenient operation and reliable effect for biological research.

Description

Feedback air-controlled pressure stress cell culture device

technical field

The invention relates to biological scientific research equipment, in particular to a feedback air-controlled pressure stress cell culture device.

Background technique

Before the invention, the compressive stress cell culture devices used in biological research are relatively expensive, and the control of continuous compressive stress is not very ideal. The effect of mechanical stress on cell growth has also been a persistent difficulty. Therefore, many laboratories are unable to carry out this research, or the cost of purchasing one is too high, or the continuous compressive stress is not ideal, resulting in inaccurate effects and affecting experimental data.

For airtight containers that are both easy to open and close and can withstand a certain amount of compressive stress (especially positive pressure), the current effects are not ideal. Present device has air inlet and air outlet more, and both are installed valves, have increased the loaded down with trivial details of device.

Contents of the invention

The purpose of the present invention is to overcome the above defects and develop a feedback air-controlled pressure stress cell culture device.

Technical scheme of the present invention is:

A feedback type air-controlled pressure stress cell culture device, its main technical feature is that the high-pressure gas storage tank is connected to the three-way pipe a through the switch, the pressure reducing valve, and the connecting pipe, and the three-way pipe a is respectively connected to the lower connecting pipe and the second connecting pipe. Conduit; the lower link conduit is connected to the micro booster pump through the syringe, the second link conduit is connected to the three-way pipe b, and the three-way pipe b is respectively connected to the third link conduit, the pressure gauge and the upper end of the sterilizing filter, and the lower end of the sterilizing filter is connected to the needle; The rubber stopper is on the measuring cylinder glass container, the upper outer reinforcing plate is on the rubber stopper, the lower outer reinforcing plate is under the cylindrical glass container, the needle penetrates the rubber stopper and enters the measuring cylinder glass container, and the cell culture dish is placed at the bottom of the measuring cylinder glass container ; The upper outer reinforcing plate and the lower outer reinforcing plate are respectively pressed on the rubber stopper and below the measuring cylinder glass container by screws and steel nails to form an airtight container; when the pressure in the airtight container reaches the pressure required for the experiment, adjust the three-way Tube a is rotated 90° counterclockwise.

The main technical feature of the feedback air-controlled pressure stress cell culture device is that the feedback control system is simulated by a micro booster pump to monitor and maintain the set pressure required for the experiment in real time.

The beneficial effect of the present invention is that the micro pressurization pump and the compressive stress cell culture device are cleverly combined to provide a compressive stress cell culture device with simple structure, convenient operation and reliable effect for biological scientific research, which greatly improves the resistance to compressive stress. Control, monitoring and maintenance, thereby improving the scientificity and accuracy of the experiment. The three-way pipe can not only connect the conduit to conduct gas, but also subtly change the direction of gas flow by rotating the valve of the three-way pipe, which not only plays the role of switch, but also plays the role of link, and simplifies the device.

Other advantages and effects of the present invention will be further explained below.

Description of drawings

Fig. 1 - schematic diagram of the structure principle of the present invention.

Figure 2 - a dynamic indication diagram of a specific application of the present invention.

The symbols in the figure represent the components as follows:

High-pressure gas storage tank 1, switch 2, pressure reducing valve 3, link conduit 4, tee pipe a5, micro pressure pump 6, syringe 7, lower link conduit 8, second link conduit 9, pressure gauge 10, third link Conduit 11, tee pipe b12, sterilizing filter 13, needle 14, screw 15, upper outer reinforcing plate 16, rubber stopper 17, measuring cylinder glass container 18, steel nail 19, cell culture dish 20, cells in the cell culture dish 21, the lower outer reinforcing plate 22.

Detailed ways

As shown in Figure 1 and Figure 2:

After the high-pressure gas storage tank 1 passes through the switch 2 on it and the pressure reducing valve 3, it is connected to the three-way pipe a5 through the link conduit 4, and the second end and the third end of the three-way pipe a5 are respectively connected to the lower link conduit 8, the second The link conduit 9; the link conduit 8 is connected to the micro pressure pump 6 through the syringe 7, the second link conduit 9 communicates with the tee pipe b12, the second end of the tee pipe b12 communicates with the pressure gauge 10 through the third link conduit 11, and the tee pipe b12 The third end is connected to the upper end of the sterilizing filter 13; the lower end of the sterilizing filter 13 is connected to the needle 14; a rubber stopper 17 is added on the measuring cylinder glass container 18 to form a closed container; the upper outer reinforcing plate 16, the lower outer reinforcing plate 22 are tightly pressed on the top of the rubber stopper 17 and the bottom of the graduated glass container 18 through four sets of screws 15 and steel nails 19; the needle 14 penetrates the rubber stopper 17 and enters the graduated glass container 18, and the cell A petri dish 20 contains cells 21 to be cultured.

The application process of the present invention is described as follows:

The high-pressure gas storage tank 1 stores a compressed mixed gas containing 5% CO2 and 95% air, simulating the gas concentration in the C02 cell culture box; after the switch 2 is opened, it is immediately closed to make the gap between the switch 2 and the pressure reducing valve 3 Store a certain amount of mixed gas; through the pressure reducing valve 3, the connecting conduit 4, the three-way pipe a5, the second connecting conduit 9, the three-way pipe b12, the sterilizing filter 13, the needle 14, the high-pressure gas storage tank 1 vector cylinder glass Load gas in the airtight container that container 18 and rubber stopper 17 form, detect the pressure in the airtight container by pressure gauge 10, after reaching the required pressure of experiment, adjust three-way pipe and be three-way pipe a5 (Fig. 2: A to B, That is, rotate 90° counterclockwise), change the flow direction of the gas, and the cell compressive stress is loaded. The use of the three-way pipe not only simplifies the device and eliminates the use of inlets, outlets and valves, but also effectively controls the flow direction of the gas, thereby effectively controlling the compressive stress required for the experiment.

Through the micro booster pump 6, the feedback control of the compressive stress of the whole device is realized. Before use, a compressive stress is set in the micro booster pump 6, and its size should be suitable with the required compressive stress of the experiment, that is, the indicated compressive stress of the pressure gauge 10, and the flow rate of the gas is set simultaneously. When the set compressive stress of the micro booster pump 6 was equivalent to the indicated compressive stress of the pressure gauge 10, the micro booster pump did not work; The pressure required for the experiment, that is, for the micro booster pump, when the actual pressure is less than the set pressure, the micro booster pump 6 will push the mixed gas in the syringe 7 at a certain flow rate until it reaches the pressure of the micro booster pump 6. Pre-set pressure. In this way, the real-time monitoring and continuous maintenance of the compressive stress required for the experiment are realized, so that the experimental conditions are more rigorous, scientific and controllable. It is a feedback control system, which not only detects the compressive stress in the experimental process in real time, but also replenishes the mixed gas in time once the compressive stress drops, until the required compressive stress of the experiment is reached and maintained.

The upper outer reinforcing plate 16, the lower outer reinforcing plate 22, four sets of screws 15, and steel nails 19 can completely seal the rubber stopper 17 and the measuring cylinder glass container 18 to form an airtight container that can withstand a certain compressive stress. Keep it for a long time without leaking. Experiments have shown that due to the fixing effect of the external reinforcement plate, the experimental pressure can be reduced by less than 1% when the pressure is kept at 100kPa for 24 hours. The needle 14 pierces through the rubber stopper 17 , and the application of compressive stress takes place via this needle 14 .

The needle 14 is connected with a sterilizing filter 13 to filter harmful substances such as impurities and bacteria in the air, and the sterilizing filter 13 is connected with a three-way pipe b12 to adjust the direction of gas in and out. The sterilizing filter 13 contains a filter membrane with a diameter of 0.22um, which can remove impurities, bacteria, mycoplasma and other harmful substances in the mixed gas in the high-pressure gas tank. The pressure reducing valve 3 adjusts the gas flow of the mixed gas, allowing the gas to slowly flow into the link guide 4, so that the pressure of the entire device increases slowly, improving the safety and controllability of the device; the micro booster pump 6 monitors the pressure in the airtight container , The micro booster pump 6 is small in size and can be placed in a C02 cell incubator. Charge the battery before use. In the C02 cell incubator, there is no need to connect to the power supply;

Description of the pressure loading process:

After opening the switch 2 of the high-pressure gas storage tank 1, close it quickly, so that a certain amount of mixed gas has accumulated between the pressure reducing valve 3 and the switch 2, and then adjust the fine-tuning of the pressure reducing valve 3 so that the mixed gas slowly flows into the connecting conduit 4 , through the three-way pipe a5, the second connecting conduit 9, the three-way pipe b12, the sterilizing filter 13, and the needle 14 into the measuring cylinder glass container 18, as shown in Figure 1; through the pressure gauge 10 on the three-way pipe b12 to detect the measuring cylinder After the pressure in the glass container 18 reaches the set pressure, rotate the three-way pipe a5 to block the entry of the mixed gas. As shown in Figure 2, a certain amount of compressed mixed gas can be stored in the glass container 18 of the graduated cylinder, thereby To achieve a certain compressive stress, pressurize the cells 21 in the cell culture dish 20 in the graduated glass container 18 .

Description of the pressure monitoring process:

In the case of Fig. 2, through the three-way pipe a5, the mixed gas changes direction (A to B, that is, rotates 90° counterclockwise) and flows into the syringe 7 of the micro booster pump 6. After reaching a certain volume, the syringe 7 will Store a certain amount of mixed gas, and then rotate the three-way pipe a5 (B to C, that is, rotate 180° clockwise or counterclockwise), so that the airflow direction of the micro booster pump 6 faces in the graduated cylinder glass container 18 to realize the compressive stress detection process.

Set the pressure in the micro booster pump 6 to be equal to the pressure required for the experiment. Once the compressive stress in the cell culture dish 20 is less than the set pressure, that is, for the micro booster pump, when the actual pressure is less than the set pressure, pressurize through the micro booster pump 6 until the set compressive stress is reached, The pressurization is stopped, so that the real-time monitoring and continuous maintenance of the compressive stress are realized.

The present invention not only detects the compressive stress in the experimental process in real time through the analog feedback control system of the micro booster pump, but also replenishes the mixed gas in time once the compressive stress drops, until the compressive stress required for the experiment is achieved and maintained, that is, real-time monitoring And maintain the set pressure required for the experiment.

Claims (1)

1. feedback-type pneumatic-control stress cell culture apparatus, is characterized in that high pressure tank is communicated with Y-tube a through switch, reducing valve, link conduit, and Y-tube a is communicated with respectively again lower link conduit, the second link conduit; Lower link conduit connects micro-force (forcing) pump by syringe, and second links conduit connecting tee pipe b, and Y-tube b the second end is communicated with tensimeter by the 3rd link conduit, and Y-tube b the 3rd end is communicated with the upper end of sterilization filter, sterilization filter lower end connection syringe needle; Rubber plug on the cylinder type Glass Containers, is upper outside gusset plate on rubber plug, and lower outside gusset plate is below the cylinder type Glass Containers, and syringe needle penetrates in rubber plug inlet cartridge type Glass Containers, cylinder type Glass Containers bottom placement Tissue Culture Dish; Upper outside gusset plate, lower outside gusset plate are oppressed on rubber plug and below the cylinder type Glass Containers by screw, steel nail respectively, form an encloses container; When the pressure in encloses container reaches the required pressure of experiment, regulate Y-tube a and be rotated counterclockwise 90 °.

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