CN118086005B

CN118086005B - Microorganism culture device

Info

Publication number: CN118086005B
Application number: CN202410481160.6A
Authority: CN
Inventors: 张俊祥; 程少丽; 王娜; 刘刚
Original assignee: Fruit Tree Institute of CAAS
Current assignee: Fruit Tree Institute of CAAS
Priority date: 2024-04-22
Filing date: 2024-04-22
Publication date: 2024-08-02
Anticipated expiration: 2044-04-22
Also published as: CN118086005A

Abstract

The present invention relates to the technical field of microorganism cultivation, and discloses a microorganism cultivation device, comprising: a main container and a plurality of subsidiary containers, the main container comprising a base, a top seat and a main wall; the main wall is configured as a wave structure; a circumferentially arranged interface is arranged on the radial outer side of the upper end surface of the top seat; the lower end surface of the top seat is configured as a conical surface; the lower end surface of the top seat is covered with a diaphragm with grooves adapted to the conical surface; the diaphragm covers the top seat so that a plurality of grooves correspond to the conical surface to define a plurality of flow channels and a distribution chamber is defined in the middle of the trough and the conical surface; each subsidiary container is provided with a first valve component, the first valve component is used to dock with the interface; the first valve component comprises a first check valve and a second check valve, the first check valve allows the fluid to flow from the flow channel to the inner cavity of the subsidiary container, the second check valve allows the fluid to flow from the inner cavity of the subsidiary container to the flow channel, the first check valve has a first preset opening pressure, and the second check valve has a second preset opening pressure.

Description

Microorganism culture device

Technical Field

The invention relates to the technical field of microorganism culture, in particular to a microorganism culture device.

Background

In the microorganism culture process, it may be necessary to transfer a portion of the microorganisms in the main culture vessel (petri dish) to the sub culture vessel for reasons including, but not limited to: based on different culture requirements, respectively transferring microorganisms in the main culture container to different auxiliary culture containers, and then adding different reagents into the different auxiliary culture containers to obtain experimental data capable of realizing comparison; the culture space as a whole can be significantly increased by transferring the microorganisms in the main culture vessel to a plurality of sub culture vessels for the purpose of increasing the number (scale) of culturing the microorganisms, so that the scale of culturing the microorganisms can be increased; for the purpose of fusing at least two separately cultured microorganisms later, specifically, first, a part of microorganisms in the main culture vessel is transferred to at least two sub culture vessels to be cultured in a different culture scheme, however, the sub culture vessels after a period of culture are fused again to continue the culture.

In the conventional transfer of microorganisms in a main culture vessel to each sub culture vessel, an open transfer method is mainly adopted, specifically, in the transfer process, it is necessary to first open the main culture vessel and/or the sub culture vessel, and then transfer microorganisms in the main culture vessel to the sub culture vessel by pouring the main culture vessel directly, or by discharging the microorganisms from an outlet at the bottom of the culture vessel, or by suction and injection using a suction and injection device. It is easy to understand that the above-described conventional means of transferring microorganisms have at least the following drawbacks: during transfer, the time and space for exposure of the microorganisms is increased, which may lead to contamination of the microorganisms; the contact possibility of operators and microorganisms is increased, and mutual pollution can be caused; the transfer operation process is complicated; the transferred sub-culture vessel needs to be separately placed on a special placement device and platform.

To overcome one or more of the drawbacks of the conventional transfer method, some patent documents provide a device for culturing microorganisms, which detachably couples a plurality of sub-culture vessels to a main culture vessel, and provides a pipeline between the main culture vessel and each sub-culture vessel, by which microorganisms in the main culture vessel can be transferred to the sub-culture vessel, so that, in theory, the device can significantly reduce the time and space for exposing microorganisms, the possibility of contacting with operators, and facilitate the integration of the culture vessels.

However, the devices for culturing microorganisms provided in these patent documents still have the following problems:

1. In some patent documents, only various transfer functions and objects of the main culture vessel and each sub culture vessel are described, and specific structures for supporting the transfer are not described in detail, for example, it is described that the main culture vessel and the sub culture vessel are arranged in an elastically compressible structure, and microorganism transfer is achieved by the pipe by compressing or rebounding the main culture vessel and the sub culture vessel, but how a control structure for controlling the on-off of the pipe is adapted to be opened or closed is not described in detail, which may cause that a structure for supporting these transfer functions cannot be obtained by a technician without performing creative work, for example, a specific structure for preventing microorganisms in the sub culture vessel from flowing back to the main culture vessel needs to be obtained with creative work during transfer, and a specific structure for achieving transfer between the sub culture vessels needs to be obtained with creative work, for example.

2. In other patent documents, devices for culturing microorganisms are provided which have large design drawbacks and which are capable of achieving fewer transfer functions. For example, each secondary culture vessel draws one flexible tube into the main culture vessel, resulting in a plurality of flexible tubes in the main culture vessel; for another example, transfer between microorganisms in each of the pair of culture vessels cannot be achieved; for another example, the transfer process causes a part of microorganisms to remain in the pipeline and not return to the main culture vessel, and for another example, reverse transfer of microorganisms in the sub culture vessel to the main culture vessel cannot be achieved.

Thus, the applicant has sought to provide a microorganism culture apparatus which is relatively rational in structural configuration and which is capable of providing more transfer functions.

Disclosure of Invention

In view of the above technical problems in the prior art, embodiments of the present invention provide a microorganism culture apparatus.

In order to solve the technical problems, the technical scheme adopted by the embodiment of the invention is as follows:

A microbial culture apparatus comprising: a main container and a plurality of sub-containers detachably attached to the main container,

The main container comprises a base, a top seat and a main wall between the base and the top seat; the main wall is configured in a wave structure to enable the main container to elastically expand and contract in an axial direction; the radial outer side of the upper end surface of the top seat is provided with circumferentially arranged interfaces, the bottoms of a plurality of auxiliary containers are detachably attached to the interfaces, and the top seat is provided with an injection port and a balance port which penetrate through the inner cavity of the main container and can be selectively opened; wherein:

The lower end surface of the top seat is configured into a conical surface; the lower end surface of the top seat is covered with a diaphragm which is adapted to the conical surface; the diaphragm is provided with a plurality of grooves which are circumferentially arranged and radially extend, and a sinking groove is formed in the middle of the diaphragm; the diaphragm covers the top seat so that a plurality of grooves and the conical surface are opposite to each other to define a plurality of flow passages, and the sinking groove and the middle part of the conical surface are opposite to each other to define a distribution cavity; the radially inner ends of the flow channels are converged and communicated to the distribution cavity, and the radially outer end of each flow channel is communicated to the interface; a hose leading from the distribution chamber to the bottom of the inner chamber of the main container;

The bottom of each auxiliary container is provided with a first valve component which is used for being in butt joint with the interface; the first valve member includes a first check valve allowing fluid to flow from the flow passage to the inner chamber of the sub-tank to be reversely shut off, and a second check valve allowing fluid to flow from the inner chamber of the sub-tank to the flow passage to be reversely shut off, and the first check valve has a first preset opening pressure, and the second check valve has a second preset opening pressure.

Preferably, the first check valve comprises a first valve sleeve, a first plunger valve core and a first spring; the first valve sleeve is fixed in a mounting hole at the bottom of the auxiliary container, a first valve cavity and a first downward step surface are defined in the first valve sleeve, a limiting table is formed at the upper end of the first plunger valve core, a second upward step surface is formed at the lower end of the first plunger valve core, and the first spring is sleeved on the first plunger valve core and is arranged between the first step surface and the second step surface; wherein:

A plurality of first valve passages which are circumferentially arranged are formed in the outer peripheral surface of the first plunger valve core, the first valve passages extend upwards from the lower end of the first plunger valve core for a section so that the first plunger valve core is provided with a first flow stopping section which is positioned at the upper part and is columnar in the outer peripheral surface, and a first flow guiding section which is positioned at the lower part and is provided with the first valve passages on the outer peripheral surface, when the first spring yields, the first plunger valve core is switched from the first flow stopping section to the first valve cavity to the first flow guiding section to be matched with the first valve cavity, and when the first flow guiding section is matched with the first valve cavity, fluid flows through the first valve cavity through the first valve passages so that the first check valve is opened;

the first plunger valve core is provided with a second valve cavity which is axially communicated, and the second check valve is arranged in the second valve cavity;

The second check valve comprises an annular valve frame and a circular valve plate; the valve frame is positioned at the upper port of the second valve cavity, and a lath is transversely arranged in the middle of the valve frame; the valve plate is positioned below the valve frame, the middle part of the valve plate is fixed on the batten, the edge of the valve plate is stopped at the edge of the valve frame, and the valve plate is configured so that the fluid in the auxiliary container can force the valve plate to elastically deform downwards with the middle part as a bending area, so that the second check valve is opened.

Preferably, a second valve member is provided between each of the first valve members and the corresponding interface; wherein:

the second valve member includes:

The second valve sleeve is arranged in the connector, the inner wall of the second valve sleeve is provided with second valve channels which are circumferentially distributed, and the second valve channels extend upwards from the lower end of the second valve sleeve for a section so that the second valve sleeve is provided with a second flow stopping section positioned at the upper part and a second flow guiding section positioned at the lower part;

A second plunger valve core disposed in the second valve housing;

a limit stop formed above the second valve sleeve for stopping the second plunger valve core;

a magnet assembly comprising a first magnet and a second magnet respectively fixed on the plunger piston and the limit spigot, wherein magnetic attraction is formed between the first magnet and the second magnet so that the plunger piston is positioned at an upper position and matched with the second flow stop section to close the interface;

the trigger frame is annular, the trigger frame is installed in the lower extreme of the first valve pocket of first check valve, the trigger frame has the conical trigger head that is located the downward bulge in middle part, after the bottom of sub-container combines in the interface, the conical trigger head pushes down to the plunger piston makes the plunger piston move down and with the cooperation of second water conservancy diversion section, thereby will the interface is opened.

Preferably, a seat is formed in the middle of the diaphragm, the seat has a valve surface facing the distribution chamber, and the upper end of the hose extends into the seat and penetrates through the valve surface; wherein:

The guide hole penetrating through the distribution cavity is formed downwards from the upper end of the top seat, the valve rod is penetrated in the guide hole, the upper end of the guide hole is screwed with the pushing nut, the lower end of the valve rod is provided with the valve head, the guide hole is internally provided with the second spring pushing against the valve rod upwards, the valve head is driven to move downwards by screwing the pushing nut so as to be abutted against the valve face to seal the upper end of the hose, and a plurality of flow channels distributed in the circumferential direction are blocked with the inner cavity of the main container so that the flow channels are communicated through the distribution cavity.

Preferably, a flushing channel is formed on the side wall of the top seat corresponding to each interface, the flushing channel penetrates through the corresponding interface, and the flushing channel is selectively opened and closed.

Preferably, a rubber stopper is removably mounted at the upper port of each of said secondary containers through which the needle assembly can pass into said secondary container.

Preferably, the secondary container is a rigid container.

Preferably, the sub-tank is an elastically stretchable tank.

Preferably, a filter is installed in the balance port.

Preferably, the top base is made of transparent plexiglas, and the main wall is made of transparent plastic material or silicone material.

Compared with the prior art, the microbial culture device provided by the embodiment of the invention has the beneficial effects that:

1. By providing the first valve part with two non-return valves, and the valve stem, microorganisms can be transferred from the main container to the sub-container, from the sub-container to the main container, from one sub-container to the other sub-container.

2. By configuring the second valve member, the interface on the primary container is automatically closed after the secondary container is separated from the primary container, thereby avoiding the external environment from affecting microorganisms in the primary container via the interface.

3. The diaphragm provided with a plurality of grooves arranged in the circumferential direction is covered on the top seat to form a plurality of flow passages for forming a passage between the auxiliary container and the main container, and the auxiliary container is communicated with the main container through the flow passages, so that the auxiliary container and the main container are prevented from being respectively communicated by a plurality of hoses.

4. The bottom of the top seat is configured with a tapered surface so that the flow channels are all inclined toward the radially inner end, which is advantageous in reducing microorganisms retained in the flow channels.

5. Other key advantages of the present invention are set forth directly and implicitly in the detailed description which follows.

An overview of various implementations or examples of the technology described in this disclosure is not a comprehensive disclosure of the full scope or all of the features of the technology disclosed.

Drawings

In the drawings, which are not necessarily drawn to scale, like numerals may describe similar components in different views. The same reference numerals with letter suffixes or different letter suffixes may represent different instances of similar components. The accompanying drawings illustrate various embodiments by way of example in general and not by way of limitation, and together with the description and claims serve to explain the inventive embodiments. Wherever possible, the same reference numbers will be used throughout the drawings to refer to the same or like parts. Such embodiments are illustrative and not intended to be exhaustive or exclusive of the present apparatus or method.

FIG. 1 is a front sectional view of a microorganism culture apparatus according to the present invention.

Fig. 2 is a plan view of the microorganism culture apparatus provided by the present invention.

Fig. 3 is a cross-sectional view taken along A-A of fig. 1.

Fig. 4 is a view showing a state in which the microorganism culture apparatus according to the present invention is used to transfer microorganisms in a main vessel to a sub-vessel.

Fig. 5 is an enlarged view of a portion B of fig. 4.

Fig. 6 is a cross-sectional view taken along the direction C-C of fig. 5.

Fig. 7 is a sectional view taken along direction D-D of fig. 5.

Fig. 8 is a view showing a state in which the microorganism culture apparatus according to the present invention is used to transfer microorganisms in the sub-tank to the main tank.

Fig. 9 is an enlarged view of a portion E of fig. 8.

FIG. 10 is a view showing a state in which a microorganism culture apparatus according to the present invention is used for transferring a microorganism from one sub-tank to another sub-tank (the sub-tank is a rigid tank).

FIG. 11 is a view showing a state in which a microorganism culture apparatus according to the present invention is used for transferring a microorganism from one sub-tank to another sub-tank (the sub-tank is an elastically stretchable and deformable tank).

Fig. 12 is an enlarged view of a portion F of fig. 10.

Reference numerals:

10-a main container; 11-footstock; 111-injection ports; 112-balancing port; 1121-a filter; 113-interface; 114-conical surface; 115-flushing the channel; 116-a guide hole; 12-a base; 13-a main wall; 14-a membrane; 20-a sub-tank; 21-a rubber bottle stopper; 31-flow channel; 32-a dispensing chamber; 33-hose; 34-a base; 341-valve face; 40-a first valve component; 50-a first non-return valve; 51-a first valve sleeve; 511-a first valve cavity; 512-a first step surface; 52-a first plunger spool; 53-a first spring; 521-a first flow stop section; 522-a first deflector segment; 523-first valve passage; 524-second step surface; 525-limit table; 526-a second valve chamber; 60-a second check valve; 61-a valve plate; 62-valve frame; 621-panels; 70-a second valve component; 71-a second valve sleeve; 711-a second stop segment; 712-a second flow directing section; 713-a second valve passage; 72-second plunger valve core; 73-limiting rabbets; 741-first magnet; 742-a second magnet; 75-triggering a frame; 81-valve stem; 811-a valve head; 82-pushing the mother; 83-a second spring; 100-syringe; 200-venting needle tube.

Detailed Description

Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs. The terms "first," "second," and the like, as used herein, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "upper", "lower", "left", "right", etc. are used merely to indicate relative positional relationships, which may also be changed when the absolute position of the object to be described is changed.

In order to keep the following description of the embodiments of the present invention clear and concise, the detailed description of known functions and known components thereof have been omitted.

The microorganism culture apparatus provided by the invention aims at providing more transfer functions for microorganisms on one hand and aims at reasonably configuring structures to realize the transfer functions on the other hand.

In the present invention, the microorganisms include, but are not limited to: microorganisms involved in agriculture, microorganisms involved in medicine, and microorganisms involved in bioengineering.

Reference to a fluid in the present invention refers to a medium containing microorganisms, and thus, reference to a fluid is to be understood as a liquid fluid rather than a gaseous fluid. The present invention does not specifically distinguish between descriptions of fluids, microorganisms, and media mixed with microorganisms, i.e., in the present invention, it is considered that these descriptions are not substantially different, and in general, a transfer fluid may be understood as a transfer microorganism, which may make the following description more concise and convenient.

As shown in fig. 1 to 12, the microorganism culture apparatus provided by the present invention comprises: a main tank 10, a plurality of sub tanks 20, and related valve members and the like mounted in the main tank 10 and the sub tanks 20.

As shown in fig. 1, the main container 10 includes: a base 12, a top base 11, and a main wall 13; the base 12 and the top base 11 are configured in a disc-shaped structure, and can be made of transparent organic glass, and the base 12 and the top base 11 can be basically considered as rigid components; the main wall 13 is interposed between the base 12 and the top base 11, and the main wall 13 may be made of transparent rubber, silica gel, and plastic having a certain elasticity, and the main wall 13 is configured in a wave structure, so that the main wall 13 can elastically stretch and retract, thereby allowing the main container 10 to elastically stretch and retract. The significant advantage of configuring the main container 10 to be elastically stretchable is that: on the one hand, when injecting or sucking the medium (fluid) mixed with the microorganisms into the main vessel 10, the main wall 13 may be expanded and contracted to avoid a large degree of variation in pressure inside the main vessel 10, thereby facilitating the injection process or the sucking process, and on the other hand, in some cases, the main vessel 10 may be enabled to press the fluid into the sub-vessel 20 attached to the main vessel 10 or suck the fluid from the sub-vessel 20 by actively compressing and stretching the main vessel 10.

The middle area of the top seat 11 is provided with an injection port 111 and a balance port 112 which penetrate through the inner cavity of the main container 10; the culture medium mixed with microorganisms can be injected into the inner cavity of the main container 10 through the injection port 111 at the beginning of the culture experiment, and after the injection is completed, the injection port 111 is blocked and closed by a rubber plug; the balance port 112 is used to balance pressure, for example, when the pressure variation in the inner cavity cannot be balanced even by autonomous expansion and contraction of the main container 10, the balance hole can be opened to communicate with the outside atmosphere to balance pressure, and preferably a filter 1121 is installed at the balance port 112 in an effort to prevent the outside air from contaminating microorganisms within the main container 10.

As shown in fig. 1 and in combination with fig. 2 to 5, the radial edge of the top seat 11 is processed with a plurality of interfaces 113 penetrating axially, and a plurality of sub-containers 20 are respectively and detachably attached to the interfaces 113 of the top seat 11; the lower end surface of the top seat 11 is configured as a conical surface 114, the conical surface 114 is a conical surface 114 with a high middle part low edge, a diaphragm 14 is attached and fixed to the lower end surface of the top seat 11, the diaphragm 14 is adapted to the conical structure of the conical surface 114 and covers the conical surface 114, a plurality of grooves which are circumferentially arranged and radially extend and sink grooves which are positioned in a middle part area are formed on the diaphragm 14, thus, the plurality of grooves of the diaphragm 14 and the conical surface 114 define a plurality of flow passages 31 which are circumferentially arranged and radially extend, the sink grooves of the diaphragm 14 and the middle part of the conical surface 114 define a distribution cavity 32, and the radially inner ends of all the grooves are converged in the middle part area and are communicated with the distribution cavity 32, and the radially outer ends of all the grooves are respectively communicated with an interface 113 of the top seat 11; a seat 34 is disposed in the middle of the diaphragm 14, a hose 33 is drawn from the seat 34 to extend to the bottom of the inner cavity of the main vessel 10, and the upper end of the hose 33 is passed through to the distribution chamber 32, so that each port 113 establishes a passage with the microorganism-mixed medium in the main vessel 10 through the corresponding flow passage 31, distribution chamber 32, and hose 33.

As shown in fig. 1 and 10 and 12, the seat 34 has a valve surface 341 facing the dispensing chamber 32, the top seat 11 corresponding to the valve surface 341 is provided with a guide hole 116 penetrating axially, the guide hole 116 is provided with a valve rod 81, a pushing nut 82 is installed above the valve rod 81, the guide hole 116 is provided with a second spring 83, the second spring 83 pushes the valve rod 81 upwards so that the pushing nut 82 is in an upper position before pushing the valve rod 81 downwards, at this time, a valve head 811 at the lower end of the valve rod 81 is retracted in the lower end of the guide hole 116, and after the valve rod 81 is forced to move downwards by screwing the pushing nut 82, the valve head 811 at the lower end of the valve rod 81 is pressed against the valve surface 341 of the seat 34, at this time, the main container 10 is blocked off from each flow passage 31 and the corresponding interface 113, and each interface 113 and flow passage 31 remain in communication through the dispensing chamber 32.

The bottom of each sub-tank 20 is provided with a first valve member 40, and after the sub-tank 20 is attached to the mouthpiece 113 of the top base 11, there is a second valve member 70 between the first valve member 40 and the mouthpiece 113, the first valve member 40 and the second valve member 70 being in abutment for controlling the flow direction of the fluid for cooperating with other components and structures to effect transfer of microorganisms between the main tank 10 and the sub-tank 20 and between the sub-tanks 20.

After the sub-tank 20 is attached to the mouthpiece 113, for example, after the sub-tank 20 is attached to the mouthpiece 113 in a screw-fit manner, the second valve member 70 causes the mouthpiece 113 to open, and after the sub-tank 20 is separated from the mouthpiece 113, the second valve member 70 closes the mouthpiece 113, thereby avoiding the mouthpiece 113 from causing the main tank 10 to communicate with the external environment without the sub-tank 20 attached.

The first valve member 40 includes a first check valve 50 and a second check valve 60; the first check valve 50 allows the fluid to flow from the flow passage 31 to the inner chamber of the sub-tank 20 to be reversely shut off, the second check valve 60 allows the fluid to flow from the inner chamber of the sub-tank 20 to the flow passage 31 to be reversely shut off, and the first check valve 50 has a first preset opening pressure and the second check valve 60 has a second preset opening pressure. Thus, as shown in fig. 4 and 5, when the pressure difference between the pressure of the inner cavity of the main vessel 10 and the pressure in the sub-vessel 20 is greater than a certain degree, that is, greater than the first preset opening pressure, the first check valve 50 is opened, and microorganisms in the main vessel 10 flow into the corresponding sub-vessel 20 through the hose 33, the distribution chamber 32, the corresponding flow passage 31, the second valve member 70, and the first check valve 50, thereby transferring microorganisms from the main vessel 10 to the sub-vessel 20; as shown in fig. 8 and 9, when the pressure difference between the pressure in the sub-tank 20 and the pressure in the main tank 10 is greater than a certain degree, that is, greater than the second preset pressure, the second check valve 60 is opened, and the microorganisms in the sub-tank 20 flow into the main tank 10 via the second check valve 60, the corresponding flow passage 31, the distribution chamber 32, and the hose 33, thereby transferring the microorganisms from the sub-tank 20 to the main tank 10.

The first and second preset pressures are set based on various pressure fluctuation factors and microorganism transfer objectives, e.g., the two preset pressures should be greater than a smaller pressure fluctuation range, because the smaller pressure fluctuation generally does not occur for transfer purposes, e.g., fluctuation in the internal cavity due to autonomous expansion or contraction within the main vessel 10, rather than for transfer purposes, e.g., pressure increase and decrease due to the occurrence of smaller gas increment or gas decrement within the sub-vessel 20, if the sub-vessel 20 is a rigid vessel of an invariable volume, rather than for transfer purposes. Thus, the values of the first preset pressure and the second preset pressure need to be set as large as possible to avoid as much as possible the occurrence of undesirable transfer situations of microorganisms. If the sub-tank 20 is a resiliently flexible tank, the first preset pressure and the second preset pressure may be configured to relatively small values, and if the sub-tank 20 is a rigid tank, the first preset pressure and the second preset pressure may be configured to relatively large values.

As shown in fig. 5, 6 and 9, the first check valve 50 and the second check valve 60 in the first valve member 40 are arranged in the following specific configurations. The first check valve 50 includes a first valve housing 51, a first plunger spool 52, and a first spring 53; the first valve sleeve 51 is fixed in a mounting hole at the bottom of the auxiliary container 20, the first valve sleeve 51 is internally provided with a first valve cavity 511 and a first step surface 512 which is positioned below the first valve cavity 511 and downward, the upper end of the first plunger valve core 52 is provided with a limit table 525 which is radially outward, the lower end of the first plunger valve core 52 is provided with a second step surface 524 which is upward, and the first spring 53 is sleeved on the first plunger valve core 52 and is arranged between the first step surface 512 and the second step surface 524; the outer peripheral surface of the first plunger valve core 52 is provided with a plurality of first valve passages 523 which are circumferentially arranged, the first valve passages 523 extend upwards from the lower end of the first plunger valve core 52 for a section so that the first plunger valve core 52 is provided with a first flow stopping section 521 which is positioned at the upper part and has a columnar surface on the outer peripheral surface and a first flow guiding section 522 which is positioned at the lower part and has the first valve passages 523 on the outer peripheral surface, when the first spring 53 yields, the first plunger valve core 52 is switched from the first flow stopping section 521 to the first flow guiding section 522 to the first valve cavity 511, and when the first flow guiding section 522 is matched with the first valve cavity 511, fluid flows through the first valve cavity 511 through the first valve passages 523, so that the first non-return valve 50 is opened; the first plunger valve core 52 is provided with a second valve cavity 526 which is axially penetrated, and the second check valve 60 is arranged in the second valve cavity 526; the second non-return valve 60 comprises an annular valve frame 62 and a circular valve plate 61; the valve frame 62 is positioned at the upper port of the second valve cavity 526, and a slat 621 is transversely arranged in the middle of the valve frame 62; the valve plate 61 is positioned below the valve frame 62 and the middle portion of the valve plate 61 is fixed to the plate 621, the edge of the valve plate 61 is stopped against the edge of the valve frame 62, and the valve plate 61 is configured such that the fluid in the sub-tank 20 can force the valve plate 61 to elastically deform downward with the middle portion as a bending region, whereby the second check valve 60 is opened. The first preset opening pressure is set by reasonably configuring the elastic coefficient and the initial compression amount of the first spring 53 such that the first spring 53 yields to open the first check valve 50 when the pressure difference between both sides of the first check valve 50 is greater than the first preset opening pressure; the second preset opening pressure is set by appropriately configuring the timing of the elastic bending deformation of the valve plate 61 in the second check valve 60 such that the valve plate 61 is bent to open the second check valve 60 when the pressure on both sides of the second check valve 60 is greater than the second preset opening pressure.

As shown in fig. 5 and 7, the second valve member 70 is configured as follows. The second valve member 70 includes: the second valve housing 71, the second plunger piston 72, the limit stop 73, the magnet assembly, and the trigger frame 75. The second valve sleeve 71 is installed in the mouthpiece 113, the inner wall of the second valve sleeve 71 is provided with second valve passages 713 arranged circumferentially, the second valve passages 713 extend upwards from the lower end of the second valve sleeve 71 for a section so that the second valve sleeve 71 has a second flow stopping section 711 at the upper part and a second flow guiding section 712 at the lower part; the second plunger valve core 72 is disposed in the second valve housing 71; a stopper notch 73 is formed above the second valve housing 71 for stopping the second plunger valve core 72; the magnet assembly comprises a first magnet 741 and a second magnet 742 respectively fixed on the plunger piston 72 and the limiting spigot 73, wherein the first magnet 741 and the second magnet 742 form magnetic attraction so that the plunger piston 72 is positioned at an upper position and is matched with the second flow stop section 711 to close the interface 113; the trigger frame 75 has a ring shape, the trigger frame 75 is mounted to the lower end of the first valve housing 51 of the first check valve 50, and the trigger frame 75 has a tapered trigger head protruding downward at the middle. Thus, after the bottom of the sub-tank 20 is coupled to the mouthpiece 113, the tapered trigger pushes down against the plunger piston 72 such that the plunger piston 72 moves down to engage with the second diversion section 712, thereby opening the mouthpiece 113 to enable microorganism transfer between the sub-tank 20 and the main tank 10 and between the sub-tank 20, while after the sub-tank 20 is separated from the main tank 10, the first magnet 741 and the second magnet 742 move up to the upper position to engage with the second diversion section 711 by magnetic attraction, thereby closing the mouthpiece 113 to prevent the external environment from contaminating microorganisms in the cavity of the main tank 10 through the mouthpiece 113.

As shown in fig. 1, the sub-tank 20 may be a rigid incompressible tank, for example, the sub-tank 20 is a plexiglass tank, and as shown in fig. 11, the sub-tank 20 may also be an elastically stretchable and deformable tank such as the main tank 10, for example, the body (or sub-wall) of the sub-tank 20 is configured in a wave structure and made of rubber, silica gel or plastic having elastic properties.

The side wall of the top seat 11 corresponding to each interface 113 is provided with a flushing channel 115, the flushing channel 115 penetrates through the corresponding interface 113, and a rubber plug is arranged at the flushing channel 115. The rubber stopper closes off the outer port of the flushing channel 115 when transferring microorganisms, and is removed when flushing the channel 115 is required, and a flushing liquid (e.g., the same liquid as the matrix in the main vessel 10) is injected into the flow channel 31 via the flushing channel 115 using a tool such as the syringe 100.

The method of using the microorganism culture apparatus described above is described below.

1. The sub-tank 20 is a rigid tank.

1) It is necessary to transfer at least part of the microorganisms in the main vessel 10 to the sub-vessel 20.

A. The sub-tanks 20 attached to the main tank 10 all require the case of microorganisms from the main tank 10.

Two ways can be used: the first way is: firstly, the rubber bottle plugs 21 at the upper ports of all the auxiliary containers 20 are opened or the rubber bottle plugs 21 are pierced by a needle component with an air exhausting function to enable the auxiliary containers 20 to be communicated with the outside atmosphere, then, the main container 10 is compressed, so that the pressure difference between the main container 10 and the bracket in the auxiliary container 20 is larger than a first preset opening pressure, the first check valves 50 at the bottoms of all the auxiliary containers 20 are opened, and microorganisms in the main container 10 sequentially pass through the hose 33, all the flow passages 31, the corresponding interfaces 113 and the corresponding first check valves 50 and enter all the auxiliary containers 20; the second way is: as shown in fig. 4, the pressure difference between the inside of the main container 10 and the inside of the sub container 20 is made to be greater than the first preset opening pressure by sequentially sucking the gas into the sub container 20 using the syringe 100, so that the microorganisms of the main container 10 sequentially flow to the sub container 20 through the corresponding flow paths 31, thereby accomplishing the transfer of the microorganisms one by one to the sub container 20. It should be noted that: if the amount of microorganisms required for all the sub-tanks 20 is large, so that the main tank 10 cannot suppress large pressure fluctuations by the autonomous expansion and contraction manner, there is a possibility that the pressure difference between the inside of the sub-tank 20 and the inside of the main tank 10 is greater than the second preset pressure during or after the transfer, and thus the microorganisms transferred into the sub-tank 20 are re-flowed to the main tank 10, the opening and closing of the balance port 112 needs to be controlled in real time for balancing the pressure.

B. the secondary containers 20 attached to the primary container 10 are only one or more, but not all, cases where microorganisms from the primary container 10 are required.

In this case, as shown in fig. 4 and 5, the microorganisms in the main vessel 10 may be transferred into the sub-vessels 20 through the corresponding flow paths 31 by sucking the gas into the corresponding sub-vessels 20 using the syringe 100. It should be noted that: if the amount of microorganisms to be transferred is large, and the second check valve 60 of the other sub-tank 20 is easily opened, so that microorganisms previously existing in the other sub-tank 20 may be transferred to the main tank 10, the opening and closing of the balance port 112 are controlled in real time.

2) The microorganisms in the sub tank 20 need to be transferred to the main tank 10.

As shown in fig. 8 and 9, the injector 100 is used to inject gas into the sub-tank 20 such that the pressure difference between the inside of the sub-tank 20 and the inside of the main tank 10 is greater than the second preset pressure, thereby forcing the second check valve 60 to open, so that microorganisms in the sub-tank 20 flow to the main tank 10 via the flow passage 31, the distribution chamber 32, and the hose 33. It should be noted that: if the amount of the transferred microorganisms is excessive and the main tank 10 cannot suppress large pressure fluctuations even by autonomous expansion and contraction, it is necessary to control the opening and closing of the balance port 112 in real time for balancing the pressure.

3) The case where microorganisms need to be transferred between the two sub-tanks 20.

First, as shown in fig. 10, the valve stem 81 is moved down by screwing the pushing nut 82 to block the valve face 341 of the seat 34 by the valve head 811, thereby cutting off the flow path 31 from the main tank 10, then the vent needle tube 200 is inserted into the sub-tank 20 for receiving microorganisms by penetrating the rubber stopper 21, the microorganisms are forced to flow through the second check valve 60 of the sub-tank 20 for administering microorganisms by the syringe 100 to the corresponding flow path 31 by injecting gas into the sub-tank 20 for administering microorganisms, then flow through the distribution chamber 32 to the flow path 31 corresponding to the sub-tank 20 for receiving microorganisms, and then enter the tank for receiving microorganisms through the corresponding first check valve 50.

2. The sub-tank 20 is an elastically stretchable and deformable tank.

As shown in fig. 11, unlike the case where the sub-tank 20 is a rigid tank, it is: instead of injecting gas by the syringe 100 by compressing the sub-tank 20, instead of sucking gas by the syringe 100 by elongating the sub-tank 20, the pressure is balanced by the characteristic of active expansion and contraction, and the pressure is balanced by the exhaust pipe.

The sub-tank 20 of this structure is more convenient and simpler to handle in transferring microorganisms than a rigid tank.

Furthermore, although exemplary embodiments have been described in the present disclosure, the scope thereof includes any and all embodiments having equivalent elements, modifications, omissions, combinations (e.g., of the various embodiments across), adaptations or alterations as would be appreciated by those in the art. The elements in the claims are to be construed broadly based on the language employed in the claims and are not limited to examples described in the present specification or during the practice of the application, which examples are to be construed as non-exclusive. It is intended, therefore, that the specification and examples be considered as exemplary only, with a true scope and spirit being indicated by the following claims and their full scope of equivalents.

The above description is intended to be illustrative and not restrictive. For example, the above-described examples (or one or more aspects thereof) may be used in combination with each other. For example, other embodiments may be used by those of ordinary skill in the art upon reading the above description. In addition, in the above detailed description, various features may be grouped together to streamline the invention. This is not to be interpreted as an intention that the disclosed features not being claimed are essential to any claim. Rather, inventive subject matter may lie in less than all features of a particular disclosed embodiment. Thus, the following claims are hereby incorporated into the detailed description as examples or embodiments, with each claim standing on its own as a separate embodiment, and it is contemplated that these embodiments may be combined with one another in various combinations or permutations. The scope of the invention should be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled.

The above embodiments are only exemplary embodiments of the present invention and are not intended to limit the present invention, the scope of which is defined by the claims. Various modifications and equivalent arrangements of this invention will occur to those skilled in the art, and are intended to be within the spirit and scope of the invention.

Claims

1.A microbial culture apparatus comprising: a main container and a plurality of sub-containers detachably attached to the main container, characterized in that,

The bottom of each auxiliary container is provided with a first valve component which is used for being in butt joint with the interface; the first valve member includes a first check valve allowing fluid to flow from the flow passage to the inner chamber of the sub-tank to be reversely shut off, and a second check valve allowing fluid to flow from the inner chamber of the sub-tank to the flow passage to be reversely shut off, and the first check valve has a first preset opening pressure, and the second check valve has a second preset opening pressure;

A seat body is formed in the middle of the diaphragm, the seat body is provided with a valve surface facing the distribution cavity, and the upper end of the hose stretches into the seat body and penetrates through the valve surface; wherein:

2. A microorganism culture apparatus according to claim 1, wherein,

The first check valve comprises a first valve sleeve, a first plunger valve core and a first spring; the first valve sleeve is fixed in a mounting hole at the bottom of the auxiliary container, a first valve cavity and a first downward step surface are defined in the first valve sleeve, a limiting table is formed at the upper end of the first plunger valve core, a second upward step surface is formed at the lower end of the first plunger valve core, and the first spring is sleeved on the first plunger valve core and is arranged between the first step surface and the second step surface; wherein:

3. The microbial cultivation apparatus according to claim 2, wherein a second valve member is provided between each of the first valve members and the corresponding interface; wherein:

the second valve member includes:

A second plunger valve core disposed in the second valve housing;

4. The microbial cultivation apparatus according to claim 1, wherein a flushing passage is provided on a side wall of the top base corresponding to each of the ports, the flushing passage penetrating to the corresponding port, the flushing passage being selectively opened and closed.

5. A microbial cultivation apparatus according to claim 1, wherein a rubber stopper is detachably mounted at the upper port of each of the sub-containers, through which the needle assembly is passed into the sub-container.

6. The microbial cultivation apparatus according to claim 1, wherein the sub-container is a rigid container.

7. The microorganism culture apparatus according to claim 1, wherein the sub-tank is an elastically stretchable tank.

8. The microbial cultivation apparatus according to claim 1, wherein a filter is installed in the balance port.

9. The microbial cultivation apparatus according to claim 1, wherein the top base is made of transparent plexiglas, and the main wall is made of transparent plastic material or silicone material.