Zebra fish double-channel timing intermittent hypoxia model experiment device

Abstract

The invention discloses a zebra fish double-channel timing intermittent hypoxia model experiment device which is characterized in that the device comprises a static environment fish tank (1) and a dynamic environment fish tank (2), upper covers (3) are arranged at top openings of the static environment fish tank (1) and the dynamic environment fish tank (2), and feeding openings are formed in the upper covers (3); a quantitative bait casting mechanism (4) is arranged at the bait casting opening, dissolved oxygen sensors (5) and temperature sensors (6) are arranged in the static environment fish tank (1) and the dynamic environment fish tank (2) respectively, the device further comprises a vacuum pump (7), the vacuum pump (7) is connected with inner cavities of the static environment fish tank (1) and the dynamic environment fish tank (2) through vacuumizing pipelines (8) respectively, and the vacuum pump (7) is connected with the vacuum pipelines (8). And an electromagnetic valve (9) is also arranged on the vacuumizing pipeline (8).

Description

Technical Field

[0001] The invention relates to an experimental device, in particular to a zebrafish dual-channel timed intermittent hypoxia model experimental device. Background Art

[0002] Sleep apnea-hypopnea syndrome (OSAHS) is a hypoventilation and / or apnea event caused by complete or partial collapse of the upper airway soft tissue during sleep. Due to repeated episodes of hypoxia and hypercapnia during sleep at night, symptoms such as repeated awakenings, fragmented sleep, decreased blood oxygen saturation, and daytime sleepiness occur. OSAHS is a high-risk factor for hypertension, coronary heart disease, diabetes, and cerebrovascular disease. It is also a common cause of traffic accidents and even sudden death at night. Therefore, OSAHS is a potentially fatal sleep breathing disorder.

[0003] Given ethical restrictions, constructing an experimental animal model of OSAHS has become the key to in-depth research on its pathogenesis. The current mainstream method is to simulate the condition of clinical OSAHS patients by having terrestrial animals intermittently inhale hypoxic gases, but this has significant defects. In contrast, zebrafish have become a better experimental subject due to their unique hypoxia-inducible factors, low cost, easy observability, and good ability to simulate the clinical OSAHS disease process. The zebrafish hypoxia model provides an important means for related research by monitoring the changes in various physiological data of embryos or adult fish in a hypoxic environment.

[0004] Existing zebrafish hypoxia culture and research model devices have significant problems when used, such as difficulty in ensuring that the internal oxygen concentration reaches the preset hypoxia standard, and failure to comprehensively evaluate the physiological responses and lung function changes of zebrafish under hypoxic conditions, which seriously affects the accuracy and reliability of the data.

[0005] Therefore, a method or device capable of solving the above problems is now needed. Summary of the invention

[0006] The present invention aims to solve the above-mentioned deficiencies in the prior art and proposes an experimental device with simple structure, ingenious design, reasonable layout, and capable of comprehensively evaluating the physiological response and lung function changes of zebrafish under hypoxic conditions.

[0007] The technical solution of the present invention is: a zebrafish dual-channel timed intermittent hypoxia model experimental device, characterized in that: the device comprises a static environment fish tank 1 and a dynamic environment fish tank 2, the top openings of the static environment fish tank 1 and the dynamic environment fish tank 2 are both provided with an upper cover 3, a feeding port is opened on the upper cover 3, a quantitative feeding mechanism 4 is provided at the feeding port, and a dissolved oxygen sensor 5 and a temperature sensor 6 are both provided in the static environment fish tank 1 and the dynamic environment fish tank 2, The device further comprises a vacuum pump 7, which is connected to the inner cavities of the static environment fish tank 1 and the dynamic environment fish tank 2 respectively through a vacuum pumping pipeline 8, and a solenoid valve 9 is also arranged on the vacuum pumping pipeline 8. The device also includes a constant temperature water tank 10, which is connected to the inner cavity of the dynamic environment fish tank 2 through an outlet pipe 12 and a return pipe 13. A water pump 14 and a flow meter 15 are arranged on the outlet pipe 12, and a water pump 14 is arranged on the return pipe 13. The connection between the return pipe 13 and the constant temperature water tank 10 is located on the bottom end surface of the constant temperature water tank 10. A biochemical cotton layer 22 and a filter screen 23 are arranged at the bottom of the constant temperature water tank 10. A liquid level sensor 11 is also provided on the inner wall of the dynamic environment fish tank 2, a camera 24 is provided on the bottom end surface of the upper cover, and an electric heating element 25 is also provided in the constant temperature water tank 10. The dissolved oxygen sensor 5 , the temperature sensor 6 , the vacuum pump 7 , the solenoid valve 9 , the liquid level sensor 11 , the water pump 14 , the flow meter 15 , the camera 24 and the electric heating element 25 are all electrically connected to the control module 16 .

[0008] The quantitative feeding mechanism 4 includes a cylindrical base 17, which is threadedly connected to the feeding port. A turntable 18 is rotatably connected to the upper part of the base 17, and a quantitative cylinder 19 that is transparent from top to bottom is arranged on the turntable 18. The horizontal section of the quantitative cylinder 19 is semicircular. A baffle 20 is arranged on the base 17 and is located below the turntable 18. A semicircular discharge port 21 is opened on the baffle 20 and matches the bottom opening of the quantitative cylinder 19.

[0009] The static environment fish tank 1 and the dynamic environment fish tank 2 are also provided with a pH sensor 26 .

[0010] Compared with the prior art, the present invention has the following advantages: This type of zebrafish dual-channel timed intermittent hypoxia model experimental device has a simple structure, ingenious design, and reasonable layout. It is designed to address the problems of traditional hypoxia experimental devices and has a special structure. It controls the vacuum pump through the control system to achieve the timed intermittent extraction of normal gas in the fish tank, thereby increasing the accuracy of the hypoxia parameters, and cooperates with the liquid level sensor to ensure the stability of the device during operation; at the same time, the dissolved oxygen sensor installed in it can monitor the changes in oxygen content in the water body in real time, and then monitor the changes and performance of zebrafish under different dissolved oxygen conditions through the camera. It not only includes a dynamic test fish tank, but also a static test fish tank for comparison. By adjusting the water change speed to simulate dynamic conditions, the comprehensiveness of the device can be effectively improved. In addition, a quantitative feeding device is set at the feeding port of the upper cover. The quantitative feeding device can ensure that the experimental process is carried out in a closed environment, ensuring the sealing and ease of operation of the entire system. This experimental device can more accurately simulate the pathological process of OSAHS, ensure the stability and accuracy of oxygen concentration during the experiment, and provide more reliable data support for the study of the pathogenesis of OSAHS and its complications. The two different experimental environments, dynamic and static, can stimulate lung function by changing the swimming speed of zebrafish to create models, thereby more comprehensively evaluating the physiological response and lung function changes of zebrafish under hypoxic conditions, improving the accuracy and practicality of the experiment.

[0011] In summary, it can be said that this experimental device has many advantages and is particularly suitable for promotion and application in this field, and its market prospects are very broad. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 2 is a top view of an embodiment of the present invention.

[0013] Figure 2 It is a front view of an embodiment of the present invention.

[0014] Figure 3 It is a three-dimensional structural schematic diagram of the quantitative feeding mechanism in an embodiment of the present invention.

[0015] Figure 4 It is a cross-sectional view of the quantitative feeding mechanism in the embodiment of the present invention.

[0016] Figure 5 Schematic diagram of the structure of the turntable part in the embodiment of the present invention.

[0017] Figure 6 Schematic diagram of the structure of the base part in the embodiment of the present invention. DETAILED DESCRIPTION

[0018] The specific implementation of the present invention will be described below with reference to the accompanying drawings. Figures 1 to 6 As shown: a zebrafish dual-channel timed intermittent hypoxia model experimental device, which includes a static environment fish tank 1 and a dynamic environment fish tank 2, the static environment fish tank 1 and the dynamic environment fish tank 2 are both provided with an upper cover 3 at the top opening, the upper cover 3 is provided with a feeding port, and a quantitative feeding mechanism 4 is provided at the feeding port, and the static environment fish tank 1 and the dynamic environment fish tank 2 are both provided with a dissolved oxygen sensor 5 and a temperature sensor 6, The device further comprises a vacuum pump 7, which is connected to the inner cavities of the static environment fish tank 1 and the dynamic environment fish tank 2 respectively through a vacuum pumping pipeline 8, and a solenoid valve 9 is also arranged on the vacuum pumping pipeline 8. The device also includes a constant temperature water tank 10, which is connected to the inner cavity of the dynamic environment fish tank 2 through an outlet pipe 12 and a return pipe 13. A water pump 14 and a flow meter 15 are arranged on the outlet pipe 12, and a water pump 14 is arranged on the return pipe 13. The connection between the return pipe 13 and the constant temperature water tank 10 is located on the bottom end surface of the constant temperature water tank 10. A biochemical cotton layer 22 and a filter screen 23 are arranged at the bottom of the constant temperature water tank 10. A liquid level sensor 11 is also provided on the inner wall of the dynamic environment fish tank 2, a camera 24 is provided on the bottom end surface of the upper cover, and an electric heating element 25 is also provided in the constant temperature water tank 10. The dissolved oxygen sensor 5 , the temperature sensor 6 , the vacuum pump 7 , the solenoid valve 9 , the liquid level sensor 11 , the water pump 14 , the flow meter 15 , the camera 24 and the electric heating element 25 are all electrically connected to the control module 16 .

[0019] The quantitative feeding mechanism 4 includes a cylindrical base 17, which is threadedly connected to the feeding port. A turntable 18 is rotatably connected to the upper part of the base 17, and a quantitative cylinder 19 that is transparent from top to bottom is arranged on the turntable 18. The horizontal section of the quantitative cylinder 19 is semicircular. A baffle 20 is arranged on the base 17 and is located below the turntable 18. A semicircular discharge port 21 is opened on the baffle 20 and matches the bottom opening of the quantitative cylinder 19.

[0020] The static environment fish tank 1 and the dynamic environment fish tank 2 are also provided with a pH sensor 26 .

[0021] The working process of the zebrafish dual-channel timed intermittent hypoxia model experimental device of the embodiment of the present invention is as follows: first, a suitable volume of water is injected into the static environment fish tank 1, the dynamic environment fish tank 2 and the constant temperature water tank 10, and then two batches of zebrafish of the same specifications and quantity are respectively placed into the static environment fish tank 1 and the dynamic environment fish tank 2, and then the upper cover 3 is buckled, at this time, the inner cavity of the static environment fish tank 1 and the dynamic environment fish tank 2 is in a closed state; The vacuum pump 7 and the electromagnetic valve 9 are controlled by the preset program in the control module 16. The vacuum pump 7 works at a certain time interval to extract the air in the static environment fish tank 1 and the dynamic environment fish tank 2, so that a certain vacuum degree is formed inside the fish tanks, and a low oxygen environment is intermittently created in the two fish tanks. During this process, the dissolved oxygen sensors 5 in the two fish tanks will monitor the dissolved oxygen content in the water in real time, and send the data changes to the control module 16 for analysis and storage; and the pH sensor 26 can monitor the pH value changes in the two fish tanks respectively, and judge the stability of the hypoxic conditions according to the pH value changes; the camera 24 is used to capture the morphological changes and vitality changes of the zebrafish in the two fish tanks in the low oxygen environment, such as whether their bodies are bent, etc.; once the collected video screen shows that the gill cover of the zebrafish has a short and irregular fluctuation, and it lasts for more than one minute, the control module will automatically stop the vacuum pump 7; During the above experiment, the control module 16 controls the water pump 14 on the outlet pipe 12 and the return pipe 13 to work, so that the water circulates in the constant temperature water tank 10 and the dynamic environment fish tank 2. On the one hand, water flow is generated in the dynamic environment fish tank 2 to stimulate the zebrafish to swim, so as to observe and collect the changes in its swimming situation in the intermittent hypoxic environment. On the other hand, since the water in the constant temperature water tank 10 is heated by the electric heating element 25, after the hot water participates in the water circulation, the water in the dynamic environment fish tank 2 is always kept at a constant temperature. The temperature sensor 6 located therein can monitor the water temperature in real time and send the water temperature data to the control module 16. In the above water changing process, the flow meter 15 can detect the flow rate of the water in real time, so as to ensure the stability of the water flow rate in the dynamic environment fish tank 2 and reduce the stress effect on the zebrafish; When it is necessary to feed, bait is placed in the quantitative cylinder 19 in the quantitative feeding mechanism 4 in advance. As long as the quantitative cylinder 19 is filled every time feeding, the quantitative feeding operation can be realized. The specific operation process is as follows. In the initial state, the quantitative cylinder 19 is located above the unopened part of the baffle 20. At this time, the bait therein will not fall. When it is necessary to feed, the operator holds the quantitative cylinder 19 and drives the turntable 18 to rotate relative to the base 17. After the quantitative cylinder 19 rotates to a position matching the semicircular discharge port 21, the bait therein will fall through the semicircular discharge port 21 to the static environment fish tank 1 or the dynamic environment fish tank 2 below, thereby realizing the quantitative feeding operation.

[0022] In the above process, various parameters in the static environment fish tank 1 and the dynamic environment fish tank 2, as well as the status and performance of the zebrafish in the intermittent hypoxic environment, are collected respectively. Through a control experiment (static environment fish tank 1), the effectiveness of the dynamic environment fish tank 2 is verified, and at the same time, differences other than the target variables (such as dissolved oxygen) in the later biological experiment process can be eliminated.

Claims

1. A zebrafish dual-channel timed intermittent hypoxia model experimental device, characterized in that: The device comprises a static environment fish tank (1) and a dynamic environment fish tank (2), wherein the static environment fish tank (1) and the dynamic environment fish tank (2) are both provided with an upper cover (3) at their top openings, a feeding port is provided on the upper cover (3), a quantitative feeding mechanism (4) is provided at the feeding port, and a dissolved oxygen sensor (5) and a temperature sensor (6) are both provided in the static environment fish tank (1) and the dynamic environment fish tank (2). The device further comprises a vacuum pump (7), wherein the vacuum pump (7) is connected to the inner cavities of the static environment fish tank (1) and the dynamic environment fish tank (2) respectively via a vacuum pumping pipeline (8), and a solenoid valve (9) is also arranged on the vacuum pumping pipeline (8). The device further comprises a constant temperature water tank (10), wherein the constant temperature water tank (10) is connected to the inner cavity of the dynamic environment fish tank (2) via a water outlet pipe (12) and a water return pipe (13), wherein a water pump (14) and a flow meter (15) are arranged on the water outlet pipe (12), and a water pump (14) is arranged on the water return pipe (13), wherein the connection between the water return pipe (13) and the constant temperature water tank (10) is located on the bottom end surface of the constant temperature water tank (10), and a biochemical cotton layer (22) and a filter screen (23) are arranged at the bottom of the constant temperature water tank (10). A liquid level sensor (11) is also provided on the inner wall of the dynamic environment fish tank (2), a camera (24) is provided on the bottom end surface of the upper cover, and an electric heating element (25) is also provided in the constant temperature water tank (10). The dissolved oxygen sensor (5), the temperature sensor (6), the vacuum pump (7), the solenoid valve (9), the liquid level sensor (11), the water pump (14), the flow meter (15), the camera (24), and the electric heating element (25) are all electrically connected to the control module (16).

2. The zebrafish dual-channel timed intermittent hypoxia model experimental device according to claim 1, characterized in that: The quantitative feeding mechanism (4) comprises a cylindrical base (17), the base (17) being threadedly connected to the feeding port, a turntable (18) being rotatably connected to the upper part of the base (17), a quantitative cylinder (19) being transparent from top to bottom being arranged on the turntable (18), the horizontal cross section of the quantitative cylinder (19) being semicircular, a baffle (20) being arranged on the base (17) and located below the turntable (18), the baffle (20) being provided with a semicircular feeding port (21) matching with the bottom opening of the quantitative cylinder (19).

3. The zebrafish dual-channel timed intermittent hypoxia model experimental device according to claim 1, characterized in that: A pH sensor (26) is also provided in the static environment fish tank (1) and the dynamic environment fish tank (2).

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