Cold spring large-scale biological in-situ fixing and culturing device

Abstract

The utility model belongs to the technical field of large-scale organism in-situ experiment equipment, and particularly relates to a cold spring large-scale organism in-situ fixing and culturing device which realizes in-situ culture of large-scale organisms through a culturing and fixing barrel, fidelity transfer is performed after fixing liquid is fixed, and sealing and cover locking of the culturing and fixing barrel are completed by utilizing a sealing cover plate, a silica gel ring and a locking hook. The in-situ experiment device is matched with a manipulator of an underwater robot, a large-scale organism sampler is used for obtaining living bodies, and after the living bodies are placed into the culture and fixing barrel, in-situ experiments of large-scale organisms are carried out; or the large organisms are extruded by a manipulator to be crushed, soaked in the fixing liquid of the culture and fixing barrel and carried to a deck position to complete fidelity transfer work of the large organisms. The size of the culture barrel and the size of the fixing barrel can be replaced according to needs, and the storage requirement is met; the locking hook and the torsion spring are used for locking the cover, so that the underwater robot is suitable for severe conditions of hybrid motion and collision, is convenient to operate and carry, and can be used for quickly and effectively culturing and fixing large biological samples.

Description

Technical Field

[0001] This invention belongs to the technical field of large-scale biological in-situ experimental equipment, specifically a large-scale cold spring biological in-situ fixation and culture device. Background Technology

[0002] Studies have shown that stress changes lead to immune stress, and conventional sampling methods cannot reproduce the normal physiological state and regulatory patterns of marine organisms. The effects of stress are even more pronounced on organisms in the symbiotic systems of the main dominant species in hydrothermal vents and cold seeps—symbiotic bacteria usually suffer irreversible loss under stress changes. How to fix the in situ RNA state of organisms under constant pressure has become an urgent task. Using large-scale biological culture and fixation methods can solve part of the problem.

[0003] However, both domestically and internationally, research on special marine life processes remains in its early stages due to insufficient in-situ detection technologies. In particular, the lack of large-scale in-situ biological culture and experimental equipment makes it impossible to characterize and verify the in-situ state of their physiological and biochemical processes. Currently, many domestic marine research institutions are developing related fixation equipment; however, most research on marine organism fixation technology focuses on large-scale biological storage facilities, which are structurally complex, have small sample sizes, and are difficult to operate. For example, utility model patent CN110763503A, published on February 7, 2020, discloses a "ROV-based in-situ fixation device for cutting large organisms such as deep-sea mussels"; utility model patent CN110763502A, also published on February 7, 2020, discloses a "ROV-based in-situ fixation device for compressing large organisms such as deep-sea mussels". Both of these fixation devices use cutting or compressing methods to break up and fix large organisms, but they cannot fix high-throughput samples. The utility model patent published on April 19, 2022, with publication number CN114371062A, discloses a "high-throughput in-situ fixation device for large deep-sea organisms based on ROV". It uses a fixation bucket to fix samples of large organisms. However, since the switch mode is touch-based, leakage is prone to occur when the lid is opened during transportation. Utility Model Content

[0004] To address the aforementioned problems with existing in-situ fixation devices, and considering the characteristics of in-situ fixation methods for large organisms such as marine mussels and marine molluscs, the purpose of this invention is to provide an in-situ fixation and cultivation device for large organisms in cold seeps. This invention utilizes the precise positioning and observation capabilities of an underwater robot to enable the capture of large organisms, and employs a novel opening and closing cover design to ensure a secure, contact-resistant seal.

[0005] The objective of this utility model is achieved through the following technical solution:

[0006] This utility model includes a cover plate, a concave groove, an upper sealing support plate, a lower sealing support plate, a locking hook, a culture and fixing tank, a sealing base, a rotating shaft pin A, a torsion spring A, a torsion spring B, a rotating shaft pin B, an opening pull rope, and a float. The bottom end of the culture and fixing tank is sealed to the sealing base, and the top end of the culture and fixing tank is fixedly connected to the interconnected upper and lower sealing support plates. A concave groove is fixedly attached to the inner surface of the tail end of the cover plate. The concave groove is hinged to one side of the upper sealing support plate via the rotating shaft pin A, and a [missing information - likely a device or component] is sleeved on the rotating shaft pin A. A torsion spring A, one end of which abuts against a concave groove, and the other end of which abuts against one side of the sealing upper support plate or the sealing lower support plate; the lower end of the locking hook is hinged to the other side of the sealing upper support plate via a rotating shaft pin B, and a torsion spring B is sleeved on the rotating shaft pin B, one end of which abuts against the locking hook, and the other end of which abuts against the other side of the sealing upper support plate or the sealing lower support plate; the float is connected to the upper end of the locking hook via an opening pull rope, and the upper end of the locking hook hooks the sealing plate when the sealing plate is closed.

[0007] Specifically: the sealing plate is equipped with an external one-way valve and an internal one-way valve. The external one-way valve allows fluid to flow from the inside to the outside. When the pressure inside the culture and fixation tank is greater than that outside, the liquid inside the culture and fixation tank flows to the outside through the external one-way valve to achieve pressure balance. The internal one-way valve allows fluid to flow from the outside to the inside. When the pressure inside the culture and fixation tank is less than that outside, the external liquid flows into the inside of the culture and fixation tank through the internal one-way valve to achieve pressure balance.

[0008] The culture and fixation tank is equipped with a sealing rubber strip along its upper edge. When the cover is opened, the sealing rubber strip helps to reduce the exchange of liquids inside and outside the culture and fixation tank.

[0009] The outer edge of the sealing rubber is provided with a plurality of film fixing holes evenly distributed along the circumferential direction. The upper edge of the culture and fixing tank is provided with bolt holes of the same number and corresponding one-to-one as the film fixing holes. The sealing rubber is fixed to the bolt holes on the upper edge of the culture and fixing tank by bolts passing through the film fixing holes. The inner side of the outer edge of the sealing rubber is divided into multiple segments along the circumferential direction.

[0010] The front end of the cover plate is provided with a cover pressure plate, and the upper end of the locking hook hooks onto the cover pressure plate when the cover plate is closed; the rear end of the cover plate is provided with a cover plate extension, and the concave groove is fixed to the inner surface of the cover plate extension; the inner surface of the cover plate is provided with a cover sealing plate, one side of the cover sealing plate is fixed to the inner surface of the cover plate, and the other side of the cover sealing plate is integrally formed with a cover sealing protrusion.

[0011] The upper and lower sealing support plates have the same shape, with a torsion spring hole and a concave groove fixing groove on one side, and a groove on the other side. A sampling hole is provided in the middle. The lower end of the locking hook is accommodated in the groove. One side of the concave opening of the groove is located in the torsion spring hole, and the other side is located in the concave groove fixing groove. A rotating shaft pin groove A is provided on the bottom surface of one side of the upper sealing support plate along the plate thickness direction. The rotating shaft pin A is accommodated in the rotating shaft pin groove A, and both ends of the rotating shaft pin A are fixed by rotating shaft pin A bolts installed on the upper sealing support plate. A rotating shaft pin groove B is provided on the bottom surface of the other side of the upper sealing support plate along the plate thickness direction. The rotating shaft pin B is accommodated in the rotating shaft pin groove B, and both ends of the rotating shaft pin B are fixed by rotating shaft pin B bolts installed on the upper sealing support plate. Both the upper and lower sealing support plates have bolt holes for interconnection.

[0012] A silicone ring, which is fixed by compression, is provided between the upper sealing plate and the lower sealing plate, and the silicone ring is fitted onto the top of the culture and fixation tank.

[0013] The concave groove has rotating shaft pin holes A on both sides of the concave opening for the rotating shaft pin A to pass through, and the bottom of the concave groove has a concave groove fixing hole for fixing to the tail end of the cover plate.

[0014] The upper end of the lock hook is hook-shaped and has a lock hook cable hole for connecting with the opening pull rope. The lower end of the lock hook has two ears, and each ear has a through hole for the rotating shaft pin B to pass through. The torsion spring B is located between the two ears.

[0015] The advantages and positive effects of this utility model are as follows:

[0016] 1. This utility model meets the in-situ fixation needs of large organisms such as mussels, molluscs, and tube worms in marine ecosystems. The main body is a mechanical structure that is less affected by the sampling depth; most of it is made of non-metallic materials with strong corrosion resistance; the culture and fixation tanks can be replaced in size as needed to meet storage requirements.

[0017] 2. This utility model is based on an underwater robot system, which can repeatedly utilize the robot's features such as precise positioning, real-time observation, and robotic arm operation to complete sample acquisition.

[0018] 3. This utility model has a simple structure and is easy to implement. It uses a locking hook and a torsion spring to complete the locking cover. Its mechanical structure is simple and suitable for mixed and collision-prone conditions. It is convenient for underwater robots to operate and carry, and can quickly and effectively cultivate and fix large biological samples. Attached Figure Description

[0019] Figure 1 This is one of the three-dimensional structural diagrams of the present invention in the open state;

[0020] Figure 2 This is the second three-dimensional structural diagram of the present invention in the open state;

[0021] Figure 3 This is a front view of the structure of this utility model in the open state;

[0022] Figure 4 for Figure 3 The left view;

[0023] Figure 5 for Figure 3 The top view from the right;

[0024] Figure 6 This is a three-dimensional structural diagram of the sealing upper support plate of this utility model;

[0025] Figure 7 This is a three-dimensional structural diagram of the sealing lower support plate of this utility model;

[0026] Figure 8 This is a three-dimensional structural diagram of the concave groove of this utility model;

[0027] Figure 9 for Figure 2 A magnified view of a section at point I;

[0028] Wherein: 1 is an external one-way valve, 2 is a cover plate, 3 is a cover pressure plate, 4 is a cover sealing plate, 5 is a cover sealing protrusion, 6 is an internal one-way valve, 7 is a cover plate extension, 8 is a concave groove, 9 is a sealing upper support plate, 10 is a sealing lower support plate, 11 is a rotating shaft pin A bolt, 12 is a locking hook, 13 is a locking hook cable hole, 14 is a rotating shaft pin B bolt, 15 is a fixing bolt A, 16 is a fixing bolt B, 17 is a culture and fixing tank, 18 is a sealing thread, 19 is a sealing base, 20 is a fixing bolt hole for the rotating position of the sealing lower support plate, 21 is a fixing bolt hole for the sealing lower support plate, 22 is... 23 is the bolt hole for fixing the upper support plate, 24 is the bolt hole for fixing the rotating position of the upper support plate, 25 is the sampling hole for the upper support plate, 26 is the sampling hole for the lower support plate, 27 is the silicone ring, 28 is the pin hole for the rotating shaft, 29 is the fixing hole for the concave groove, 30 is the torsion spring hole, 31 is the fixing groove for the concave groove, 32 is the pin for the rotating shaft, 33 is the sealing rubber, 34 is the fixing hole for the film, 35 is the upper edge of the culture and fixing tank, 36 is the torsion spring B, 37 is the pin for the rotating shaft B, 38 is the opening pull rope, 39 is the float, 40 is the bolt hole, 41 is the groove, and 42 is the pin groove for the rotating shaft B. Detailed Implementation

[0029] The present invention will now be described in further detail with reference to the accompanying drawings.

[0030] like Figures 1-5As shown, this utility model includes a cover plate 2, a concave groove 8, an upper sealing support plate 9, a lower sealing support plate 10, a locking hook 12, a culture and fixing tank 17, a sealing base 19, a rotating shaft pin A32, a torsion spring A, a torsion spring B36, a rotating shaft pin B37, an opening pull rope 38, and a float 39. The bottom end of the culture and fixing tank 17 is connected to the sealing base 19 via a sealing thread 18, providing a sealing function. The top end of the culture and fixing tank 17 is fixedly connected to the interconnected upper sealing support plate 9 and lower sealing support plate 10. A concave groove 8 is fixedly attached to the inner surface of the tail end of the cover plate 2, and the concave groove 8 is connected to the upper sealing support plate 9 via a rotating shaft pin A32. The device is side-hinged, with a torsion spring A fitted on the rotating shaft pin A32. One end of the torsion spring A abuts against the concave groove 8, and the other end of the torsion spring A abuts against one side of the sealing upper support plate 9 or the sealing lower support plate 10. The lower end of the locking hook 12 is hinged to the other side of the sealing upper support plate 9 via the rotating shaft pin B37. A torsion spring B36 is fitted on the rotating shaft pin B37. One end of the torsion spring B36 abuts against the locking hook 12, and the other end of the torsion spring B36 abuts against the other side of the sealing upper support plate 9 or the sealing lower support plate 10. The float ball 39 is connected to the upper end of the locking hook 12 via the opening pull rope 38. The upper end of the locking hook 12 hooks the sealing plate 2 when the sealing plate 2 is closed.

[0031] In this embodiment, the front end of the cover plate 2 is provided with a cover pressure plate 3, and the upper end of the locking hook 12 hooks the cover pressure plate 3 when the cover plate 2 is closed; the rear end of the cover plate 2 is provided with a cover plate extension 7, and the concave groove 8 is fixed to the inner surface of the cover plate extension 7; the inner surface of the cover plate 2 is provided with a cover sealing plate 4, one side of the cover sealing plate 4 is fixed to the inner surface of the cover plate 2 by multiple fixing bolts A15, and the other side of the cover sealing plate 4 is integrally formed with a cover sealing protrusion 5. After the cover plate 2 is closed, the cover sealing plate 4 contacts the sealing upper support plate 9, and the cover sealing protrusion 5 squeezes the silicone ring 27 to complete the sealing effect, thus playing a sealing role.

[0032] In this embodiment, an external one-way valve 1 and an internal one-way valve 6 are respectively installed on the cover plate 2. The external one-way valve 1 is threaded onto the cover plate 2, and the fluid flow direction of the external one-way valve 1 is from the inside to the outside. When the internal pressure of the culture and fixation tank 17 is greater than that of the outside, the liquid inside the culture and fixation tank 17 flows to the outside through the external one-way valve 1, completing the pressure balance. In this embodiment, the internal one-way valve 6 is threaded onto the cover plate 2, and the liquid flow direction of the internal one-way valve 6 is from the outside to the inside. When the internal pressure of the culture and fixation tank 17 is less than that of the outside, the external liquid flows into the interior of the culture and fixation tank 17 through the internal one-way valve 6, completing the pressure balance.

[0033] like Figures 1-7As shown, in this embodiment, the sealing upper support plate 9 and the sealing lower support plate 10 have the same shape. One side of the sealing upper support plate 9 has a torsion spring hole 30 and a concave groove fixing groove 31, while the other side has a groove 41. An upper support plate sampling hole 25 is located in the middle of the sealing upper support plate 9. Similarly, one side of the sealing lower support plate 10 has a torsion spring hole 30 and a concave groove fixing groove 31, while the other side has a groove 41. A lower support plate sampling hole 26 is located in the middle of the sealing lower support plate 10. The upper support plate sampling hole 25 and the lower support plate sampling hole 26 are glued to the outer top surface of the culture and fixing tank 17 using acrylic adhesive.

[0034] The lower end of the locking hook 12 is accommodated in the groove 41 on the other side of the sealing upper support plate 9 and the sealing lower support plate 10. The torsion spring A is located in the torsion spring hole 30 on one side of the sealing upper support plate 9 and the sealing lower support plate 10. A rotating shaft pin groove A24 is formed on the bottom surface of one side of the sealing upper support plate 9 along the plate thickness direction. The rotating shaft pin A32 is accommodated in the rotating shaft pin groove A24. The two ends of the rotating shaft pin A32 are fixed by rotating shaft pin A bolts 11 installed on the sealing upper support plate 9. A rotating shaft pin groove B42 is formed on the bottom surface of the other side of the sealing upper support plate 9 along the plate thickness direction. The rotating shaft pin B37 is accommodated in the rotating shaft pin groove B42. The two ends of the rotating shaft pin B37 are fixed by rotating shaft pin B bolts 14 installed on the sealing upper support plate 9. The upper sealing plate 9 has multiple upper sealing plate fixing bolt holes 22 around the sampling hole 25 in the circumferential direction, and multiple upper sealing plate rotation fixing bolt holes 23 on one side of the upper sealing plate 9. The lower sealing plate 10 has the same number of lower sealing plate fixing bolt holes 21 around the sampling hole 26 in the circumferential direction as the upper sealing plate fixing bolt holes 22, and on one side of the lower sealing plate 10 has the same number of lower sealing plate rotation fixing bolt holes 23, and corresponding lower sealing plate rotation fixing bolt holes 20. The upper sealing plate 9 and the lower sealing plate 10 are fixed together by bolts through the corresponding upper sealing plate fixing bolt holes 22, lower sealing plate fixing bolt holes 21, and corresponding upper sealing plate rotation fixing bolt holes 23 and lower sealing plate rotation fixing bolt holes 20. A silicone ring 27, which is fixed by compression, is provided between the lower surface of the upper sealing plate 9 and the upper surface of the lower sealing plate 10. The silicone ring 27 is fitted onto the top of the culture and fixation tank 17.

[0035] In this embodiment, a sealing rubber sheet 33 is installed on the upper edge 35 of the culture and fixation tank 17. Multiple film fixing holes 34 are evenly distributed along the circumferential direction on the outer edge of the sealing rubber sheet 33. Bolt holes 40, corresponding to the number of film fixing holes 34, are evenly distributed along the circumferential direction on the upper edge 35 of the culture and fixation tank. The sealing rubber sheet 33 is fixed to the bolt holes 40 on the upper edge 35 of the culture and fixation tank by bolts passing through the film fixing holes 34. The inner side of the outer edge of the sealing rubber sheet 33 is divided into multiple petals along the circumferential direction; in this embodiment, it has eight petals. When the cover plate 2 is opened, the sealing rubber sheet 33 facilitates the exchange of liquids inside and outside the culture and fixation tank 17.

[0036] like Figures 1-8 As shown, in this embodiment, the concave opening of the concave groove 8 has rotating pin holes A28 on both sides for the rotating pin A32 to pass through. A concave groove fixing hole 29 is provided at the bottom of the concave groove 8. The concave groove 8 is fixed to the inner surface of the rear extension 7 of the cover plate by a fixing bolt 16 passing through the concave groove fixing hole 29. One side of the concave opening of the concave groove 8 is located in the torsion spring hole 30 on the upper sealing plate 9 and the lower sealing plate 10, and the other side of the concave opening of the concave groove 8 is located in the concave groove fixing groove 31 on the upper sealing plate 9 and the lower sealing plate 10. The rear extension 7 of the cover plate, the concave groove 8, the upper sealing plate 9, and the lower sealing plate 10 work together to complete the rotation action. The concave groove 8, utilizing the combined action of the torsion spring A and the rotating pin A32, possesses torsional kinetic energy to complete the rotation that opens the cover plate 2. The locking hook 12 utilizes the combined action of the torsion spring B36 and the rotating shaft pin B37 to possess torsional kinetic energy. The sealing plate 3 works in conjunction with the locking hook 12 when the sealing plate 2 is closed to complete the locking action. The float 39, through its pulling action, opens the pull rope 38 and the locking hook cable hole 13, ultimately causing the locking hook 12 to move back and forth, completing the opening and closing action.

[0037] like Figures 1-9 As shown, in this embodiment, the upper end of the locking hook 36 is hook-shaped and has a locking hook cable hole 13 for connecting with the opening pull rope 38. The lower end of the locking hook 36 has two ears with a gap between them. Each ear has a through hole for the rotating shaft pin B37 to pass through. The torsion spring B36 is located between the two ears.

[0038] This utility model relates to a method for in-situ fixation and culture of large cold seep organisms, comprising the following steps:

[0039] Step A: Overall disassembly and cleaning of the shore-based end. The large-scale in-situ fixation and culture device for cold seeps is disassembled and then cleaned.

[0040] Step B: After cleaning, the components are assembled. In the initial state, the culture medium or fixative is stored in the culture and fixation tank 17. Press the cover plate 2 to rotate the concave tank 8. The cover plate 2 is closed by rotating the pivot pin A32. The upper end of the locking hook 12 hooks onto the cover plate 2 to complete the locking state. During the closing process of the cover plate 2, the liquid in the culture and fixation tank 17 will form a positive pressure state, and the pressure will be released through the external one-way valve 1.

[0041] Step C: The large-scale cold seep organism in-situ fixation and culture device is placed in the carrier of the underwater robot. The sealed base 19 has a set area size and is carried to the seabed by the underwater robot. During the carrying process, it may roll or collide. Due to the function of the locking hook 12, the liquid inside the culture and fixation tank 17 can be well protected and prevented from leaking.

[0042] Step D: After reaching the seabed, use the underwater robot's manipulator to lift the float 39, which will unlock the locking hook 12 by moving it back and forth. The torsion of the torsion spring A will open the cover plate 2. Use the live sampling device or the underwater robot's manipulator to grab the large creature. During the grabbing process, the live sampling device can protect the large creature well, but the underwater robot's manipulator will directly grab it and damage the shell or skin of the large creature, causing it to break.

[0043] Step E: Using a live sampling device or the robotic arm of an underwater robot, a large organism is placed through the eight-lobed sealing rubber 8 into a culture and fixation tank 17. The large organism is then immersed in the culture solution or fixation solution inside the culture and fixation tank 17 to cultivate or fix it.

[0044] Step F is repeated, followed by step E, to obtain multiple types of organisms. Then, the underwater robot's manipulator presses the cover plate 2, causing the concave groove 8 to rotate. The cover plate 2 is closed by rotating the pivot pin A32. The upper end of the locking hook 12 hooks onto the cover pressure plate 3 to complete the locking state. During the closing process of the cover plate 2, the water in the cultivation and fixing tank 17 will form a positive pressure state, which will be released through the external one-way valve 1.

[0045] Step G, the biological cultivation process, requires a long time; depending on the experimental needs, it is placed on the seabed for the experiment; the biological fixation process requires less time and can be brought back at an opportune time.

[0046] Step H involves using the robotic arm of an underwater robot to place the large-scale cold seep organism in-situ fixation and cultivation device into the robot's carrier. The sealed base 19 has a predetermined area size, and the device is carried to the deck position. During transport, it may roll or collide, but the locking hook 12 effectively protects the liquid inside the cultivation and fixation tank 17, preventing leakage.

[0047] Step 1: Use an underwater robot to bring back a large-scale in-situ fixation and culture device for cold seeps;

[0048] Step J: After recycling, manually open the sealing plate 2 to obtain the sample.

[0049] This invention enables in-situ culture of large organisms using a culture and fixation tank 17, followed by faithful transport after fixation with a fixative solution. The culture and fixation tank 17 is sealed and locked using a capping plate 2, a silicone ring 27, and a locking hook 12. In conjunction with an underwater robotic arm, a large organism sampler is used to obtain live organisms, which are then placed in the culture and fixation tank 17 for in-situ experiments. Alternatively, the robotic arm can be used to crush the large organisms, immerse them in the fixative solution within the culture and fixation tank 17, and then transport them to the deck for faithful transport.

Claims

1. A cold spring macrobenthos in-situ fixation and culture device, characterized by: The cover plate (2), the concave groove (8), the sealing upper supporting plate (9), the sealing lower supporting plate (10), the lock hook (12), the culture and fixing barrel (17), the sealing base (19), the rotating shaft pin A (32), the torsion spring A, the torsion spring B (36), the rotating shaft pin B (37), the opening pull rope (38) and the floating ball (39) are connected, wherein the bottom end of the culture and fixing barrel (17) is sealingly connected with the sealing base (19), the top end of the culture and fixing barrel (17) is fixedly connected with the sealing upper supporting plate (9) and the sealing lower supporting plate (10) which are connected with each other, the inner surface of the tail end of the cover plate (2) is fixedly connected with the concave groove (8), the concave groove (8) is hingedly connected with one side of the sealing upper supporting plate (9) through the rotating shaft pin A (32), the rotating shaft pin A (32) is sleeved with the torsion spring A, one end of the torsion spring A abuts against the concave groove (8), the other end of the torsion spring A abuts against one side of the sealing upper supporting plate (9) or one side of the sealing lower supporting plate (10), the lower end of the lock hook (12) is hingedly connected with the other side of the sealing upper supporting plate (9) through the rotating shaft pin B (37), the rotating shaft pin B (37) is sleeved with the torsion spring B (36), one end of the torsion spring B (36) abuts against the lock hook (12), the other end of the torsion spring B (36) abuts against the other side of the sealing upper supporting plate (9) or the other side of the sealing lower supporting plate (10), the floating ball (39) is connected with the upper end of the lock hook (12) through the opening pull rope (38), and the upper end of the lock hook (12) hooks the cover plate (2) when the cover plate (2) is closed.

2. The cold spring macro-biota in-situ fixation and culture apparatus according to claim 1, characterized in that: The cover plate (2) is respectively provided with the external one-way valve (1) and the built-in one-way valve (6), the fluid flow direction of the external one-way valve (1) is from inside to outside, when the internal pressure of the culture and fixing barrel (17) is greater than the external pressure, the liquid in the culture and fixing barrel (17) flows to the outside through the external one-way valve (1), and the pressure balance is completed, the liquid flow direction of the built-in one-way valve (6) is from outside to inside, when the internal pressure of the culture and fixing barrel (17) is less than the external pressure, the external liquid flows into the inside of the culture and fixing barrel (17) through the built-in one-way valve (6), and the pressure balance is completed.

3. The cold spring macro-biota in-situ fixation and culture apparatus according to claim 1, characterized in that: The sealing rubber (33) is arranged on the culture and fixing barrel upper edge (35) of the culture and fixing barrel (17), when the cover plate (2) is opened, the liquid exchange between the inside and the outside of the culture and fixing barrel (17) is relieved through the sealing rubber (33).

4. The cold spring macro-biota in-situ immobilizing and culturing device according to claim 3, characterized in that: A plurality of rubber sheet fixing holes (34) are uniformly arranged on the outer edge of the sealing rubber (33) in the circumferential direction, a plurality of bolt holes (40) corresponding to the rubber sheet fixing holes (34) are uniformly arranged on the culture and fixing barrel upper edge (35) in the circumferential direction, the sealing rubber (33) is fixed through the bolt holes (34) and the bolt holes (40) on the culture and fixing barrel upper edge (35), and the inner side of the outer edge of the sealing rubber (33) is divided into multiple petals in the circumferential direction.

5. The cold spring macro-biota in-situ immobilization and cultivation apparatus according to claim 1, characterized in that: The front end of the cover plate (2) is provided with a cover pressing plate (3), and the upper end of the lock hook (12) hooks the cover pressing plate (3) when the cover plate (2) is closed; the tail end of the cover plate (2) is provided with a cover plate rear extension (7), and the concave groove body (8) is fixed to the inner surface of the cover plate rear extension (7); the inner surface of the cover plate (2) is provided with a cover sealing plate (4), one side of the cover sealing plate (4) is fixed to the inner surface of the cover plate (2), and the other side of the cover sealing plate (4) is integrally formed with a cover sealing protrusion (5).

6. The cold spring macro-biota in-situ immobilization and cultivation apparatus according to claim 1, characterized in that: The sealing upper supporting plate (9) and the sealing lower supporting plate (10) are the same shape, one side of each is provided with a torsion spring hole (30) and a concave groove body fixing groove (31), the other side is provided with a recess (41), and a sampling hole is provided in the middle, the lower end of the lock hook (12) is accommodated in the recess (41), and one side of the concave groove body (8) is located in the torsion spring hole (30) and the other side is located in the concave groove body fixing groove (31); the bottom surface of one side of the sealing upper supporting plate (9) is provided with a rotating shaft pin groove A (24) in the thickness direction of the plate, the rotating shaft pin A (32) is accommodated in the rotating shaft pin groove A (24), and the two ends of the rotating shaft pin A (32) are packaged and fixed through the rotating shaft pin A bolt (11) installed on the sealing upper supporting plate (9); the bottom surface of the other side of the sealing upper supporting plate (9) is provided with a rotating shaft pin groove B (42) in the thickness direction of the plate, the rotating shaft pin B (37) is accommodated in the rotating shaft pin groove B (42), and the two ends of the rotating shaft pin B (37) are packaged and fixed through the rotating shaft pin B bolt (14) installed on the sealing upper supporting plate (9); the sealing upper supporting plate (9) and the sealing lower supporting plate (10) are both provided with bolt holes for mutual connection.

7. The cold spring macro-biota in-situ immobilization and cultivation apparatus according to claim 1, characterized in that: The sealing upper supporting plate (9) and the sealing lower supporting plate (10) are provided with a silica gel ring (27) fixed by extrusion force between them, and the silica gel ring (27) is sleeved on the top end of the culture and fixing barrel (17).

8. The cold spring macro-biota in-situ immobilization and cultivation apparatus according to claim 1, characterized in that: The two sides of the concave groove body (8) are provided with rotating shaft pin holes A (28) for the rotating shaft pin A (32) to pass through, and the bottom of the concave groove body (8) is provided with a concave groove body fixing hole (29) for fixing with the tail end of the cover plate (2).

9. The cold spring macro-biota in-situ immobilization and cultivation apparatus of claim 1, wherein: The upper end of the lock hook (12) is hook-shaped and is provided with a lock hook cable hole (13) for connecting with an opening pull rope (38), the lower end of the lock hook (12) is provided with double ears, through holes are provided in the double ears for the rotating shaft pin B (37) to pass through after the double ears, and the torsion spring B (36) is located between the double ears.

Images