CN115232713A - Portable laboratory simple and easy device of oxygen in getting rid of culture medium - Google Patents

Abstract

The invention discloses a portable laboratory simple device for removing oxygen in a culture medium, which comprises a gas pipe with a partial pressure valve connected to the outlet of the top of an oxygen-free gas cylinder; a first bottle mouth is arranged on the top of a first liquid culture medium storage bottle and the second bottle mouth, the first conveying pipe, the first exhaust pipe and the second conveying pipe all run through the first bottle mouth and are sealedly connected with the first bottle mouth, and the third conveying pipe runs through the second bottle mouth and is connected with the second bottle mouth. The mouth is sealed and connected; the second liquid culture medium storage bottle is provided with a third bottle mouth and a fourth bottle mouth on the top, and the fourth conveying pipe, the second exhaust pipe and the fifth conveying pipe all pass through the third bottle mouth and are connected with the third bottle mouth. The bottle mouth is sealed and connected, the fourth conveying pipe is connected with the second conveying pipe, the fifth conveying pipe is connected with the third conveying pipe, and the sixth conveying pipe is sealed and connected with the fourth bottle mouth; the first conveying pipe is connected with the first conveying pipe through the first branch pipe The exhaust pipe is connected. The device of the invention has the advantages of small volume, simple operation and low price.

Description

A portable laboratory simple device for removing oxygen from culture medium

technical field

The invention relates to the technical field of microbial experimental equipment, in particular to a portable laboratory simple device for removing oxygen in a culture medium.

Background technique

Anaerobic microorganisms are widely distributed in the natural environment, including water, soil, and sediments. There are many types of anaerobic microorganisms, including denitrifying bacteria, fermentation bacteria, methanogens, etc., which not only drive the biogeochemical cycle of carbon and nitrogen elements in the environment, but also are closely related to the production of greenhouse gases in the atmosphere. The preparation of anaerobic microbial liquid medium is the key to culturing anaerobic microorganisms.

At present, the preparation of anaerobic microbial liquid culture medium generally adopts the cooked meat medium method, pyrogallic acid method, vacuum method, anaerobic tank culture method, and anaerobic glove box method.

Among them, the principle of the cooked meat medium method is to use the unsaturated fatty acids and sulfhydryl groups contained in the meat residue in the medium to absorb the oxygen in the medium, thereby reducing the redox potential, and covering the liquid surface with a layer of Vaseline or liquid paraffin can be used to isolate the air and form a good anaerobic condition to meet the needs of the growth of anaerobic bacteria. However, this method can only be applied to the cultivation of specific anaerobic microorganisms.

The principle of the pyrogallic acid method is to use pyrogallic acid to absorb a large amount of oxygen quickly after being added to an alkaline solution to form a dark brown pyrogallic acid orange, which can effectively absorb oxygen in any closed container. Conditions conducive to the growth of anaerobic bacteria. However, it will produce a small amount of carbon monoxide during the oxidation process, which will poison microorganisms to a certain extent, so the growth of some CO-sensitive anaerobic bacteria will be inhibited.

The principle of the vacuum method is to discharge the medium and the gas in the headspace of the container through the vacuum pumping system to realize the replacement of oxygen and other anaerobic others, thereby completing the preparation of the anaerobic medium. However, this method is time-consuming and difficult to remove oxygen from the liquid medium to the greatest extent possible.

The principle of the anaerobic tank method is to remove the oxygen in the closed container through the oxygen-removing particles, thereby providing an anaerobic environment for the growth of anaerobic microorganisms. However, it cannot realize the anaerobic environment of medium preparation and microbial inoculation, and can only ensure the anaerobic environment of the cultivation process.

The principle of the anaerobic glove box method: it is made of transparent rigid plastic into an anaerobic glove box with good sealing performance. The whole process is operated and handled in the box by the rubber gloves on the box, so that the culture is not in contact with the air, and the anaerobic environment is always maintained. However, its instruments are expensive and bulky, and are not suitable for the cultivation of extreme anaerobic microorganisms (such as seabed bacteria and deep soil bacteria).

SUMMARY OF THE INVENTION

The purpose of the present invention is to overcome the problems in the prior art, to provide a portable device that does not rely on expensive anaerobic glove boxes and vacuum pumps, the device is small in size, simple in operation and low in price, and can be freely operated to remove oxygen in the culture medium. Laboratory simple devices are of great significance.

The invention provides a portable laboratory simple device for removing oxygen in a culture medium, comprising: an oxygen-free gas cylinder, an air delivery pipe with a partial pressure valve is connected to the top outlet; a first liquid culture medium storage bottle, the top of which is provided with a second A bottle mouth and a second bottle mouth, a first conveying pipe with a first valve, a first exhaust pipe with a second valve and a third valve, and a second conveying pipe with a fourth valve all pass through the first bottle mouth And it is sealed with the first bottle mouth, wherein the top of the first delivery pipe is communicated with the gas delivery pipe, the bottom ends of the first delivery pipe and the second delivery pipe are all immersed in the liquid culture medium, and the third delivery pipe with the fifth valve Passing through the second bottle mouth and sealingly connected with the second bottle mouth, the top of the third delivery pipe is located above the second bottle mouth, and the bottom end of the third delivery pipe is immersed in the liquid medium; the second liquid medium storage bottle, the top A third bottle mouth and a fourth bottle mouth are provided, the fourth conveying pipe with the sixth valve, the second exhaust pipe with the seventh valve and the fifth conveying pipe with the eighth valve all pass through the third bottle mouth and are connected with each other. The third bottle mouth is sealed and connected, the top end of the fourth conveying pipe located above the third bottle mouth is connected with the top end of the second conveying pipe, the top end of the fifth conveying pipe is connected with the top end of the third conveying pipe, and the fourth conveying pipe is connected to the top end of the third conveying pipe. The bottom ends of the five conveying pipes are all immersed in the liquid culture medium, and the sixth conveying pipe with the ninth valve is tightly connected with the fourth bottle mouth; The valve communicates with the first exhaust pipe between the valve and the third valve.

Preferably, the second conveying pipe is communicated with the first conveying pipe through the second branch pipe with the eleventh valve, and the fourth conveying pipe is connected with the third branch pipe with the twelfth valve and is located under the seventh valve. The second exhaust pipe communicates.

Preferably, a first sealing cap is detachably connected to the first bottle mouth, and the first sealing cap is sealed and connected with the first conveying pipe, the first exhaust pipe and the second conveying pipe respectively, and the second bottle A second sealing cap is detachably connected to the mouth, the second sealing cap is sealingly connected to the third conveying pipe, and a third sealing cap is detachably connected to the third bottle mouth, and the third sealing cap is respectively connected to the fourth conveying tube. The pipe, the second exhaust pipe and the fifth conveying pipe are sealedly connected, and a fourth sealing cover is detachably connected to the fourth bottle mouth, and the fourth sealing cover is sealedly connected with the sixth conveying pipe.

Preferably, both the first sealing cover and the third sealing cover include a first cover body and three pagoda-shaped sealing pipes arranged at the bottom of the first cover body and sealingly connected with the first cover body. The pipe, the first exhaust pipe and the second conveying pipe or the fourth conveying pipe, the second exhaust pipe and the fifth conveying pipe respectively pass through the three pagoda-shaped sealing pipes, and the second sealing cover and the fourth sealing cover both include The second cover body and a pagoda-shaped connecting pipe arranged at the bottom of the second cover body and sealingly connected with the second cover body, the third conveying pipe or the sixth conveying pipe respectively penetrates the connecting pipe.

Preferably, the bottoms of the first sealing cover, the second sealing cover, the third sealing cover and the fourth sealing cover are all provided with O-rings.

Preferably, the volume fraction ratio of nitrogen to carbon dioxide in the oxygen-free gas cylinder is 80:20.

Preferably, the pressure of the partial pressure valve is 0.1-0.2 MPa.

Preferably, the first liquid culture medium storage bottle and the second liquid culture medium storage bottle are made of glass material, the gas delivery pipe is a rigid PU pipe, the first delivery pipe and the first exhaust pipe are made of glass. , the second delivery pipe, the third delivery pipe, the second exhaust pipe, the fourth delivery pipe, the first branch pipe, the second branch pipe, and the third branch pipe are all silicone hoses, and the pagoda-shaped sealing pipe and the pagoda-shaped connecting pipe All are made of stainless steel.

Preferably, the first conveying pipe and the gas conveying pipe, the fourth conveying pipe and the second conveying pipe, and the fifth conveying pipe and the third conveying pipe can be detachably connected.

Preferably, the volume of the first liquid culture medium storage bottle is 1L-5L, and the volume of the second liquid culture medium storage bottle is 50mL-200mL.

Compared with the prior art, the beneficial effects of the present invention are:

1. The present invention can realize aeration and deoxygenation of the liquid culture medium in the first liquid culture medium storage bottle. Specific operation: Open the first valve, the second valve and the third valve, so that the oxygen-free gas enters the liquid culture medium of the first liquid culture medium storage bottle through the air delivery pipe and the first delivery pipe, so that the liquid culture medium originally existing in the liquid culture medium will be removed. And the oxygen in the headspace is released by the input oxygen-free gas through the first exhaust pipe, and finally achieves the purpose of aeration and deoxygenation of the culture medium.

2. The present invention can realize aeration and deoxygenation and anaerobic sub-packaging of the liquid medium in the second liquid medium storage bottle. Specific operation: by opening the tenth valve, the third valve, the fourth valve, the sixth valve and the seventh valve; the oxygen-free gas enters the first liquid through the gas transmission pipe, the first transmission pipe, the first branch pipe and the first exhaust pipe The headspace of the medium storage bottle liquid; the first liquid medium storage bottle medium flows into the second liquid medium storage bottle through the second conveying pipe and the fourth conveying pipe under the pressure of the gas; along with the second liquid culture The volume of the medium in the base storage bottle is gradually increased, and the original oxygen in the second liquid medium storage bottle is then discharged through the second exhaust pipe. When the volume of the medium in the second liquid medium storage bottle reaches the maximum, open the eleventh valve, the twelfth valve, the eighth valve, the fifth valve, the second valve and the third valve on the fifth delivery pipe, The oxygen-free gas enters the headspace of the second liquid culture medium storage bottle through the gas delivery pipe, the first delivery pipe, the second branch pipe, the second delivery pipe, the fourth delivery pipe, the third branch pipe and the second exhaust pipe; Under pressure, the liquid culture medium in it flows back into the first liquid culture medium storage bottle through the fifth conveying pipe and the third conveying pipe, and the gas in the headspace of the first liquid culture medium storage bottle passes through the first exhaust with the pressure of the returning liquid. The tube is drained; when the volume of the medium in the second liquid medium storage bottle is about to reach the target mark, the anaerobic aliquot of the medium is completed.

3. The anaerobic microbial culture medium preparation device provided by the present invention does not rely on expensive anaerobic glove boxes and vacuum pumps. The whole device is small in size, simple in operation, low in price, and free to operate; in addition, the device can be combined with cysteine Strict anaerobic media can be formulated with oxygen scavengers.

Description of drawings

Fig. 1 is the overall structure schematic diagram of the present invention;

Fig. 2 is the structural representation of the first liquid culture medium storage bottle of the present invention;

Fig. 3 is the structural representation of the second liquid culture medium storage bottle of the present invention;

4 is a schematic structural diagram of the first sealing cover and the third sealing cover of the present invention (where a is a bottom view; b is a longitudinal cross-sectional view);

5 is a schematic structural diagram of the second sealing cover and the fourth sealing cover of the present invention (where a is a bottom view; b is a longitudinal cross-sectional view).

Description of reference numbers:

1. Oxygen-free gas cylinder, 2. Gas delivery pipe, 3. First liquid culture medium storage bottle, 4. First delivery pipe, 5. First exhaust pipe, 6. Second delivery pipe, 7. Third delivery pipe , 8. The second liquid culture medium storage bottle, 9. The fourth delivery pipe, 10. The second exhaust pipe, 11. The fifth delivery pipe, 12. The sixth delivery pipe, 13. The first branch pipe, 14. The second Branch pipe, 15. Third branch pipe, 16. First cover body, 17. Sealing pipe, 18. Second cover body, 19. Connecting pipe, 20. O-ring seal.

Detailed ways

The specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings 1-5, but it should be understood that the protection scope of the present invention is not limited by the specific embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present invention.

Example

This embodiment provides a portable laboratory simple device for removing oxygen in a culture medium, and achieves the purpose of the present invention through basic necessary technical features.

Specifically, as shown in Figures 1-5, a portable laboratory simple device for removing oxygen in a culture medium provided by the present invention includes: an oxygen-free gas cylinder 1, a first liquid culture medium storage bottle 3 and a second liquid culture The base storage bottle 8, wherein the top outlet of the oxygen-free gas cylinder 1 is connected with an air delivery pipe 2 with a partial pressure valve; the top of the first liquid culture medium storage bottle 3 is provided with a first bottle mouth and a second bottle mouth, with a The first conveying pipe 4 with a valve, the first exhaust pipe 5 with the second valve and the third valve, and the second conveying pipe 6 with the fourth valve all pass through the first bottle mouth and are tightly connected with the first bottle mouth , wherein the top of the first delivery pipe 4 is communicated with the gas delivery pipe 2, the bottom ends of the first delivery pipe 4 and the second delivery pipe 6 are all immersed in the liquid culture medium, and the third delivery pipe 7 with the fifth valve runs through the second The bottle mouth is sealed with the second bottle mouth, the top of the third delivery pipe 7 is located above the second bottle mouth, and the bottom end of the third delivery pipe 7 is immersed in the liquid culture medium; the top of the second liquid culture medium storage bottle 8 The third bottle mouth and the fourth bottle mouth are provided, the fourth conveying pipe 9 with the sixth valve, the second exhaust pipe 10 with the seventh valve and the fifth conveying pipe 11 with the eighth valve all run through the third bottle The mouth is sealed and connected with the third bottle mouth, the top of the fourth conveying pipe 9 located above the third bottle mouth is connected with the top of the second conveying pipe 6, and the top of the fifth conveying pipe 11 is communicated with the top of the third conveying pipe 7. , the bottom ends of the fourth conveying pipe 9 and the fifth conveying pipe 11 are all immersed in the liquid culture medium, and the sixth conveying pipe 12 with the ninth valve is tightly connected with the fourth bottle mouth; The first branch pipe 13 of the valve communicates with the first exhaust pipe 5 located between the second valve and the belt third valve.

The second conveying pipe 6 communicates with the first conveying pipe 4 through the second branch pipe 14 with the eleventh valve, and the fourth conveying pipe 9 is located below the seventh valve through the third branch pipe 15 with the twelfth valve. The second exhaust pipe 10 is connected.

Specifically, a first sealing cover is detachably connected to the first bottle mouth, and the first sealing cover is sealed with the first conveying pipe 4, the first exhaust pipe 5 and the second conveying pipe 6 respectively. A second sealing cap is detachably connected to the two bottle mouths, the second sealing cap is sealingly connected to the third conveying pipe 7, and a third sealing cap is detachably connected to the third bottle mouth, and the third sealing caps are respectively connected with The fourth conveying pipe 9 , the second exhaust pipe 10 and the fifth conveying pipe 11 are sealed and connected, the fourth bottle mouth is detachably connected with a fourth sealing cover, and the fourth sealing cover is sealed with the sixth conveying pipe 12 .

Specifically, both the first sealing cover and the third sealing cover include a first cover body 16 and three pagoda-shaped sealing tubes 17 arranged at the bottom of the first cover body 16 and sealingly connected to the first cover body 16 . The first conveying pipe 4 , the first exhaust pipe 5 and the second conveying pipe 6 or the fourth conveying pipe 9 , the second exhaust pipe 10 and the fifth conveying pipe 11 respectively pass through the three pagoda-shaped sealing pipes 17 . Both the second sealing cover and the fourth sealing cover include a second cover body 18 and a pagoda-shaped connecting pipe 19 arranged at the bottom of the second cover body 18 and sealed with the second cover body 18 . The six conveying pipes 12 respectively pass through the connecting pipes 19 .

Specifically, the bottoms of the first sealing cover, the second sealing cover, the third sealing cover and the fourth sealing cover are all provided with an O-ring 20 .

Specifically, the volume fraction ratio of nitrogen to carbon dioxide in the oxygen-free gas cylinder 1 is 80:20.

Specifically, the pressure of the partial pressure valve is 0.1 MPa.

Specifically, the first liquid culture medium storage bottle 3 and the second liquid culture medium storage bottle 8 are both made of glass material, the gas delivery tube 2 is a rigid PU tube, the first delivery tube 4, the first The exhaust pipe 5, the second conveying pipe 6, the third conveying pipe 9, the second exhaust pipe 10, the fourth conveying pipe 12, the first branch pipe 13, the second branch pipe 14 and the third branch pipe 15 are all silicone hoses, Both the pagoda-shaped sealing tube and the pagoda-shaped connecting tube are made of stainless steel.

Specifically, the first conveying pipe 4 and the gas conveying pipe 2 and the fourth conveying pipe 9 and the second conveying pipe 6 can be detachably connected.

Specifically, the volume of the first liquid medium storage bottle 3 is 1L-5L, and the volume of the second liquid medium storage bottle 8 is 50mL-200mL.

The use method of the portable laboratory simple device for removing oxygen in the culture medium:

(1) aeration and deaeration of liquid culture medium in the first liquid culture medium storage bottle 3:

1. Put the liquid medium into the first liquid medium storage bottle 3, and then build a simple laboratory device for removing oxygen in the medium as shown in Figure 1.

2. Open the main valve of the oxygen-free gas cylinder 1, and adjust the air pressure on the valve to 0.1-0.2MPa; at the same time, open the first valve on the first conveying pipe 4, the second valve on the first exhaust pipe 5 and the first valve on the first exhaust pipe 5 respectively. Three valves, and keep other valves closed. At this time, the gas in the oxygen-free gas cylinder 1 enters the bottom of the liquid culture medium in the first liquid culture medium storage bottle 3 through the gas delivery pipe 2 and the first delivery pipe 4, which originally existed in the liquid culture medium. Oxygen in the base and headspace is input and oxygen-free gas is released through the first exhaust pipe 5 .

3. After operating the aeration for 1 hour as in 2, close all valves, adjust the pressure of the partial pressure valve to 0, and close the main valve of the oxygen-free gas cylinder 1, so that the medium in the first liquid culture medium storage bottle 3 can be stored. Efficient oxygen removal.

(2) Sub-packaging of culture medium:

1. After completing (1) part of the operation, keep the main valve of the oxygen-free gas cylinder 1 open and close other valves; open the tenth valve on the first branch pipe 13, the third valve on the first exhaust pipe 5, the second valve on the second The fourth valve on the delivery pipe 6 , the sixth valve on the fourth delivery pipe 9 , and the seventh valve on the second exhaust pipe 10 .

2. The gas output from the oxygen-free gas cylinder 1 enters the headspace of the first liquid culture medium storage bottle 3 through the gas pipe 2, the first transport pipe 4, the first branch pipe 13 and the first exhaust pipe 5 in turn; The continuous increase in the pressure of the body causes the liquid medium of the first liquid medium storage bottle 3 to flow into the second liquid medium storage bottle 8 through the second delivery pipe 6 and the fourth delivery pipe 9, and at the same time, the liquid medium originally present in the second liquid medium storage bottle 8 The gas in the culture medium storage bottle 8 is discharged through the second exhaust pipe 10 by the flowing liquid.

3. When the medium in the second liquid medium storage bottle 8 reaches the maximum capacity, it means that all the gas originally existing in the headspace of the second liquid medium storage bottle 8 is exhausted, so that the second liquid medium storage bottle can be removed. All headspace oxygen in bottle 8.

4. Keep the main valve of the oxygen-free gas cylinder 1 open, and close other valves at the same time; open the eleventh valve on the second branch pipe 14, the twelfth valve on the third branch pipe 15, and the fifth delivery pipe 11 in turn. Eight valves, the fifth valve on the third delivery pipe 7, the second valve (top) and the third valve (bottom) on the first exhaust pipe 5.

5. The gas in the oxygen-free gas cylinder 1 passes through the gas delivery pipe 2, via the gas delivery pipe 2, the first delivery pipe 4, the second branch pipe 14, the second delivery pipe 6, the fourth delivery pipe 9, the third branch pipe 15 and the second The exhaust pipe 10 enters the headspace of the second liquid culture medium storage bottle 8; thereafter, due to the pressure of the headspace gas of the second liquid medium storage bottle 8, the liquid medium therein is transported through the fifth delivery pipe 11 and the third delivery The pipe 7 flows back into the first liquid culture medium storage bottle 3 , and at the same time, the gas in the headspace of the first liquid culture medium storage bottle 3 is released through the first exhaust pipe 5 due to the increase of the liquid volume. When the remaining volume of the second liquid medium storage bottle 8 drops to the target volume (for example, the 50mL second liquid medium storage bottle 8 should be filled with 20-30mL of medium; the 100mL second liquid medium storage bottle 8 should be filled with 40 mL of medium). ~60mL medium; 200mL second liquid medium storage bottle 8 should be filled with 100~140mL medium), close all valves; in this way, anaerobic subpackaging of liquid medium can be realized.

6. Then disconnect the connection between the fourth conveying pipe 9 and the second conveying pipe 6, the fifth conveying pipe 11 and the third conveying pipe 7.

7. The culture medium in the second liquid culture medium storage bottle 8 can be combined with a cysteine scavenger (0.5 g/L cysteine hydrochloride combined with 0.002 g/L resazurin) to prepare a strict anaerobic medium. Among them, cysteine is a reducing agent, which can remove traces of oxygen in the liquid; resazurin is an oxygen concentration indicator. When the oxygen indicator resazurin is colorless in the medium, it can be considered that the medium is strict Anaerobic state; when resazurin appears pink in the medium, the medium is in a microaerobic state; when resazurin appears blue in the medium, the medium is in an aerobic state. Regarding the operation of adding the cysteine oxygen scavenger to the medium of the second liquid medium storage bottle 8, refer to (3) the step of bacterial inoculation.

(3) Bacterial inoculation:

1. Use a sterile anaerobic syringe to absorb the bacterial liquid to be inoculated, and remove the gas in the liquid as much as possible through the movement of the piston in the syringe;

2. Remove the needle of the syringe, communicate the syringe with the sixth delivery pipe 12, open the ninth valve on the sixth delivery pipe 12, and transfer the bacterial liquid in the syringe to the second liquid culture medium storage bottle 8;

3. Close the ninth valve on the sixth delivery pipe 12, and pull out the syringe.

(4) Collection of liquid or gas samples in culture bottles

1. Place the second liquid culture medium storage bottle 8 upright (for gas sample collection) or upside down (for liquid sample collection);

2. Connect the syringe with the sixth delivery tube 12 with a sterile and anoxic syringe with the needle removed, open the ninth valve on the sixth delivery tube 12, and use the syringe to absorb the gas or bottle in the headspace of the second liquid culture medium storage bottle 8 liquid in;

3. After the end, place the second liquid culture medium storage bottle 8 upright, close the ninth valve on the sixth delivery pipe 12, and pull out the syringe.

Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, and substitutions can be made in these embodiments without departing from the principle and spirit of the invention and modifications, the scope of the present invention is defined by the appended claims and their equivalents.

Claims (10)

1. A portable laboratory simplified apparatus for removing oxygen from a culture medium, comprising:

the oxygen-free gas steel cylinder (1) is connected with a gas pipe (2) with a partial pressure valve at the top outlet;

the liquid culture medium storage bottle comprises a first liquid culture medium storage bottle (3), wherein a first bottle opening and a second bottle opening are formed in the top of the first liquid culture medium storage bottle, a first conveying pipe (4) with a first valve, a first exhaust pipe (5) with a second valve, a first exhaust pipe (5) with a third valve and a second conveying pipe (6) with a fourth valve penetrate through the first bottle opening and are in sealing connection with the first bottle opening, the top end of the first conveying pipe (4) is communicated with an air conveying pipe (2), the bottom ends of the first conveying pipe (4) and the second conveying pipe (6) are immersed in a liquid culture medium, a third conveying pipe (7) with a fifth valve penetrates through the second bottle opening and is in sealing connection with the second bottle opening, the top end of the third conveying pipe (7) is located above the second bottle opening, and the bottom end of the third conveying pipe (7) is immersed in the liquid culture medium;

a second liquid culture medium storage bottle (8), wherein a third bottle opening and a fourth bottle opening are arranged at the top of the second liquid culture medium storage bottle, a fourth conveying pipe (9) with a sixth valve, a second exhaust pipe (10) with a seventh valve and a fifth conveying pipe (11) with an eighth valve penetrate through the third bottle opening and are in sealing connection with the third bottle opening, the top end of the fourth conveying pipe (9) above the third bottle opening is connected with the top end of the second conveying pipe (6), the top end of the fifth conveying pipe (11) is communicated with the top end of the third conveying pipe (7), the bottom ends of the fourth conveying pipe (9) and the fifth conveying pipe (11) are immersed in a liquid culture medium, and a sixth conveying pipe (12) with a ninth valve is in sealing connection with the fourth bottle opening;

the first conveying pipe (4) is communicated with a first exhaust pipe (5) positioned between the second valve and the third valve through a first branch pipe (13) with a tenth valve.

2. A portable laboratory facility for the removal of oxygen from culture media according to claim 1, wherein said second delivery pipe (6) is connected to said first delivery pipe (4) through a second branch pipe (14) with an eleventh valve, and said fourth delivery pipe (9) is connected to a second exhaust pipe (10) located below said seventh valve through a third branch pipe (15) with a twelfth valve.

3. The portable simple laboratory device for removing oxygen from a culture medium according to claim 2, wherein a first sealing cover is detachably connected to the first bottle opening, the first sealing cover is respectively connected with the first delivery pipe (4), the first exhaust pipe (5) and the second delivery pipe (6) in a sealing manner, a second sealing cover is detachably connected to the second bottle opening, the second sealing cover is connected with the third delivery pipe (7) in a sealing manner, a third sealing cover is detachably connected to the third bottle opening, the third sealing cover is respectively connected with the fourth delivery pipe (9), the second exhaust pipe (10) and the fifth delivery pipe (11) in a sealing manner, a fourth sealing cover is detachably connected to the fourth bottle opening, and the fourth sealing cover is connected with the sixth delivery pipe (12) in a sealing manner.

4. The laboratory simple apparatus for portable oxygen removal from a culture medium according to claim 3, wherein the first sealing cover and the third sealing cover each comprise a first cover (16) and three pagoda-shaped sealing tubes (17) disposed at the bottom of the first cover (16) and hermetically connected to the first cover (16), the first delivery tube (4), the first exhaust tube (5), the second delivery tube (6) or the fourth delivery tube (9), the second exhaust tube (10), and the fifth delivery tube (11) respectively extend through the three pagoda-shaped sealing tubes (17), the second sealing cover and the fourth sealing cover each comprise a second cover (18) and a pagoda-shaped connecting tube (19) disposed at the bottom of the second cover (18) and hermetically connected to the second cover (18), and the third delivery tube (7) or the sixth delivery tube (12) respectively extend through the connecting tubes (19).

5. The laboratory simple device for removing oxygen from culture medium according to claim 3, wherein the first sealing cover, the second sealing cover, the third sealing cover and the fourth sealing cover are provided with O-shaped sealing rings (20) at the bottom.

6. The laboratory mounted device for portable removal of oxygen from culture medium according to claim 1, wherein the volume fraction ratio of nitrogen to carbon dioxide in said oxygen-free gas cylinder (1) is 80.

7. The laboratory simplified apparatus for removing oxygen from a culture medium according to claim 1, wherein the pressure of the partial pressure valve is 0.1-0.2MPa.

8. The simple laboratory device for portable removal of oxygen from a culture medium according to claim 4, wherein the first liquid culture medium storage bottle (3) and the second liquid culture medium storage bottle (8) are made of glass material, the gas pipe (2) is a hard PU pipe, the first conveying pipe (4), the first exhaust pipe (5), the second conveying pipe (6), the third conveying pipe (9), the second exhaust pipe (10), the fourth conveying pipe (12), the first branch pipe (13), the second branch pipe (14) and the third branch pipe (15) are silica gel hoses, and the pagoda-shaped sealing pipe and the pagoda-shaped connecting pipe are made of stainless steel material.

9. The laboratory simple device for portable removal of oxygen from culture medium according to claim 3, wherein the first delivery pipe (4) and the gas pipe (2), the fourth delivery pipe (9) and the second delivery pipe (6), and the fifth delivery pipe (11) and the third delivery pipe (7) are detachably connected.

10. The laboratory simplified apparatus for portable removal of oxygen from a culture medium as claimed in claim 1, wherein the first liquid culture medium storage bottle (3) has a volume of 1L to 5L, and the second liquid culture medium storage bottle (8) has a volume of 50mL to 200mL.

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