High-throughput zebra fish medicine screening device and screening method

Abstract

The invention belongs to the technical field of zebra fish medicine screening, and discloses a high-flux zebra fish medicine screening device which comprises a box body, an optical imaging system is arranged under the inner portion of the box body, a plurality of supporting rods are symmetrically arranged on the two sides of the optical imaging system, and automatic motion control systems are arranged at the tops of the supporting rods on the two sides. A high-flux structure unit is arranged at the center of the bottom of the automatic motion control system, the top of the automatic motion control system is connected with an automatic medicine conveying system, and a temperature control system is further arranged in the box body. According to the device, constant-temperature control inside the box body is achieved through the temperature control system, gradient establishment of different concentrations of drugs is achieved through cooperation of the automatic motion control system and the automatic drug conveying system, finally motion behavior images of juvenile zebrafish are collected through the optical imaging system, and the experiment efficiency is greatly improved. The device is high in overall automation level, short in experimental period, simple in manual operation and low in labor cost.

Description

Technical Field

[0001] The present invention relates to the technical field of zebrafish drug screening, in particular to a high-throughput zebrafish drug screening device and a screening method. Background Art

[0002] Zebrafish have important applications in drug screening and disease research. Their physiological and genetic characteristics are highly similar to those of humans, including the conservation of gene sequences, gene functions associated with human diseases, and similar organ structures and developmental processes. The zebrafish's transparency, rapid reproduction, and ease of manipulation in biological research make it an ideal model for drug development and drug screening. Drug screening using zebrafish can evaluate the efficacy and toxicity of a large number of compounds in a short period of time, providing valuable data and insights for drug development. Therefore, the development of high-throughput zebrafish drug screening devices and methods can significantly improve the efficiency and accuracy of drug screening, promoting the discovery of new drugs and their translation into clinical applications.

[0003] The currently reported technologies and systems for zebrafish drug screening are mainly divided into two categories: 1. Evaluating drug properties by constructing a zebrafish drug screening model. For example, Chinese patent CN111388684A (publication date July 10, 2020) discloses a method for evaluating the efficacy of drugs against Alzheimer's disease using transgenic zebrafish. A zebrafish disease model is constructed and cultured in a specific drug concentration solution, and the drug properties are detected by analyzing the behavioral trajectory of the juvenile fish. This method can quickly and efficiently evaluate the effects of anti-Alzheimer's drugs, but the types of drugs are very limited, and the overall level of automation of the experiment is low. When faced with high-throughput experimental samples, the experimental cycle is very long. 2. Provide technical means for high-throughput zebrafish drug screening through the development of new experimental equipment. For example, Chinese patent CN205964520U (publication date February 22, 2017) discloses a culture vessel for high-throughput zebrafish drug screening. The device can prevent drug mixing errors and improve work efficiency, but this experimental device is very dependent on manual operation and has a low level of automation. For another example, Chinese patent CN204848875U (publication date December 9, 2015) discloses an auxiliary device for zebrafish drug screening. The device includes a cell culture plate with a scale. Adding culture water and drugs according to the scale can ensure the accuracy of drug concentration, improve the accuracy of zebrafish screening experiments, speed up the experiment and improve the experimental efficiency. However, the experimental device is also very dependent on manual operation and cannot provide technical support for high-throughput zebrafish drug screening.

[0004] The above-mentioned technologies and systems for zebrafish drug screening provide novel technical methods and experimental platforms for zebrafish drug screening. However, there are common problems such as the need for a large degree of manual operation and intervention, huge labor costs, extremely low automation levels, and long experimental cycles. These problems are often even more difficult to achieve when faced with high-throughput experimental samples. Summary of the Invention

[0005] The purpose of the present invention is to provide a high-throughput zebrafish drug screening device and screening method to solve the problems raised in the above background technology.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] A device for high-throughput zebrafish drug screening comprises a housing, an optical imaging system disposed directly below the housing, a plurality of support rods symmetrically disposed on either side of the optical imaging system, an automated motion control system disposed on top of each support rod, a high-throughput structural unit disposed at the center of the bottom of the automated motion control system, an automated drug delivery system connected to the top of the automated motion control system, and a temperature control system disposed within the housing.

[0008] The automated motion control system includes a mounting base provided at the top of the support rods on both sides, a single motion shaft being longitudinally provided on the top of the mounting base, two surfaces of the two longitudinally provided single motion shafts being connected to the bottom of the horizontally provided single motion shafts via motion connection structures, and the surfaces of the horizontally provided single motion shafts being connected to an automated drug delivery system via motion connection structures;

[0009] The high-throughput structural unit includes a high-throughput platform, a plurality of porous plates are evenly arranged on the surface of the high-throughput platform, and a plate taking slot is symmetrically arranged on both sides of each porous plate;

[0010] The automated drug delivery system includes an automated drug delivery system shell arranged on the outside of the motion connection structure on the laterally arranged moving single-axis surface. The bottom end of the interior of the automated drug delivery system shell is symmetrically connected to a delivery power device through the main structure of the automated drug delivery system. A plurality of drug test tubes are connected to the sides of the two delivery power devices that are away from each other.

[0011] Further preferably, a stepper motor is provided at one end of each of the longitudinally arranged and transversely arranged single motion axes, and each of the stepper motors is connected to a position positioning switch.

[0012] Further preferably, a plurality of connection holes are symmetrically provided on both sides of the high-throughput platform, and the connection holes are connected to the surface of the mounting seat at the top of the support rod in conjunction with fastening bolts.

[0013] Further preferably, a drug taking groove is provided at the position of the drug test tube on the top of the automatic drug delivery system housing, and the automatic drug delivery system housing and the laterally arranged motion connection structure are fixedly connected by positioning holes and fastening bolts.

[0014] Further preferably, the delivery power device includes a power box body connected to the top of the main structure of the automated drug delivery system, a micropump control circuit board is arranged inside the power box body, one end of the micropump control circuit board is connected to a liquid micropump, one end of the liquid micropump is provided with a liquid inlet connected to the drug test tube on the adjacent side, and the other end of the liquid micropump is provided with a liquid outlet, both of the liquid micropumps are connected to the instillation needle arranged at the bottom center of the main structure of the automated drug delivery system through the liquid outlet and the delivery tube, and the other end of the micropump control circuit board is provided with a delivery power device power interface and a delivery power device control interface.

[0015] Further preferably, the temperature control system includes a hot air interface arranged on the rear side of the box, a heater is connected to the outside of the hot air interface, a fan is also arranged between the hot air interface and the heater, and a temperature sensor is also arranged inside the box, and the temperature sensor is electrically connected to the temperature controller.

[0016] Further preferably, the optical imaging system includes an area light source arranged directly below the interior of the box, and a plurality of cameras are evenly arranged on the surface of the area light source corresponding to the center position of the multi-well plate.

[0017] Further preferably, the entire box body is made of a frame structure and an acrylic plate.

[0018] The present invention also provides a screening method for a high-throughput zebrafish drug screening device, comprising the following steps:

[0019] Step 1: Turn on the temperature control system to ensure that the internal temperature of the box is at a constant temperature of 28°C;

[0020] Step 2: Add zebrafish larvae to the wells on the surface of the multi-well plate, and place the multi-well plate containing the zebrafish larvae in the high-throughput structure unit, and wait for 30 minutes to allow the zebrafish larvae to adapt to the internal environment of the box 1;

[0021] Step 3: Prepare multiple drugs to be tested, and add the multiple drugs to be tested into the drug test tubes through the drug taking slot;

[0022] Step 4: The automated drug delivery system is driven to move by the automated motion control system. When the dropping needle provided at the bottom center of the main structure of the automated drug delivery system moves to the position directly above the corresponding hole on the surface of the multi-well plate, the drug to be tested is dropped into the hole on the surface of the multi-well plate by the automated drug delivery system.

[0023] Step 5: Capture images of the zebrafish larvae's movement behavior through an optical imaging system and collect the data;

[0024] Step 6: Conduct behavioral analysis on the collected zebrafish larvae movement behavior images.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The device uses a temperature control system to maintain constant temperature within the chamber. It also collaborates with an automated motion control system and an automated drug delivery system to establish gradients of varying drug concentrations. Finally, an optical imaging system captures images of zebrafish larvae's movement behavior, significantly improving experimental efficiency. The device boasts a high level of automation, short experimental cycles, simple manual operation, and low labor costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0028] Figure 2 It is a structural schematic diagram of the automatic motion control system of the present invention;

[0029] Figure 3 Schematic diagram of the structure of the optical imaging system of the present invention;

[0030] Figure 4 Schematic diagram of the structure of the high-throughput structural unit of the present invention;

[0031] Figure 5 Schematic diagram of the structure of the automated drug delivery system of the present invention;

[0032] Figure 6 It is a structural schematic diagram of the conveying power device of the present invention;

[0033] Figure 7 is a schematic diagram of a temperature control system of the present invention;

[0034] In the figure: 1. Box; 2. Automated motion control system; 3. Optical imaging system; 4. High-throughput structural unit; 5. Automated drug delivery system; 6. Support rod; 7. Frame structure; 8. Hot air interface; 9. Stepper motor; 10. Positioning switch; 11. Motion connection structure; 12. Motion single axis; 13. Area light source; 14. Camera; 15. Multi-well plate; 16. Connection hole; 17. High-throughput platform; 18. Well plate picking slot; 19. Drug picking slot; 20. Delivery power unit; 21. Automated drug delivery system housing; 22. Drug test tube; 23. Positioning hole; 24. Automated drug delivery system main structure; 25. Delivery power unit power interface; 26. Delivery power unit control interface; 27. Temperature control system; 28. Heater; 29. Fan; 30. Temperature sensor; 31. Temperature controller; 211. Liquid outlet; 222. Liquid inlet; 233. Liquid micropump; 244. Micropump control circuit board. DETAILED DESCRIPTION

[0035] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0036] See also Figure 1-Figure 7 , the present invention provides a technical solution:

[0037] A high-throughput zebrafish drug screening device includes a housing 1, an optical imaging system 3 is disposed directly below the housing 1, a plurality of support rods 6 are symmetrically disposed on both sides of the optical imaging system 3, an automated motion control system 2 is disposed on top of the support rods 6 on both sides, a high-throughput structural unit 4 is disposed at the bottom center of the automated motion control system 2, an automated drug delivery system 5 is connected to the top of the automated motion control system 2, and a temperature control system 27 is also disposed inside the housing 1;

[0038] The automated motion control system 2 includes a mounting base provided at the top of the support rods 6 on both sides, a motion single shaft 12 being longitudinally provided on the top of the mounting base, and the surfaces of the two longitudinally provided motion single shafts 12 are connected to the bottom of the transversely provided motion single shaft 12 via motion connection structures 11, and the surfaces of the transversely provided motion single shafts 12 are connected to the automated drug delivery system 5 via the motion connection structures 11;

[0039] The high-throughput structural unit 4 includes a high-throughput platform 17, on the surface of which a plurality of porous plates 15 are evenly arranged, and each porous plate 15 is symmetrically provided with a plate taking slot 18 on both sides.

[0040] The automated drug delivery system 5 includes an automated drug delivery system shell 21 arranged on the outside of the motion connection structure 11 on the surface of the laterally arranged motion single shaft 12. The bottom end of the interior of the automated drug delivery system shell 21 is symmetrically connected to a delivery power device 20 through an automated drug delivery system main structure 24. A plurality of drug test tubes 22 are connected to the sides of the two delivery power devices 20 that are away from each other.

[0041] In the present invention, a stepper motor 9 is provided at one end of each of the longitudinally arranged and transversely arranged moving single shafts 12, and each stepper motor 9 is respectively connected to a position positioning switch 10. The model of the stepper motor 9 is 5756.

[0042] In the present invention, a plurality of connection holes 16 are symmetrically provided on both sides of the high-throughput platform 17 , and the connection holes 16 are connected to the surface of the mounting seat at the top of the support rod 6 with fastening bolts.

[0043] In the present invention, a drug taking groove 19 is provided at the top of the automated drug delivery system housing 21 corresponding to the position of the drug test tube 22, and the automated drug delivery system housing 21 and the laterally arranged motion connection structure 11 are fixedly connected through positioning holes 23 and fastening bolts.

[0044] In the present invention, the delivery power unit 20 includes a power box connected to the top of the automated drug delivery system main structure 24. A micropump control circuit board 244 is disposed within the power box. A liquid micropump 233 is connected to one end of the micropump control circuit board 244. One end of the liquid micropump 233 is provided with a liquid inlet 222 connected to the adjacent drug test tube 22. The other end of the liquid micropump 233 is provided with a liquid outlet 211. Both liquid micropumps 233 are connected to a dropper located at the bottom center of the automated drug delivery system main structure 24 through the liquid outlet 211 and the delivery tube. The other end of the micropump control circuit board 244 is provided with a delivery power unit power interface 25 and a delivery power unit control interface 26. The drug test tubes 22 are eight 15ml centrifuge tubes, four on each side. The centrifuge tube covers have through-holes that accommodate the extraction tube and the liquid micropump 233 for liquid delivery.

[0045] In the present invention, the temperature control system 27 includes a hot air interface 8 provided on the rear side of the box body 1, a heater 28 is connected to the outside of the hot air interface 8, a fan 29 is provided between the hot air interface 8 and the heater 28, and a temperature sensor 30 is provided inside the box body 1, and the temperature sensor 30 is electrically connected to the temperature controller 31. The heater 28 and the fan 29 are an integrated structure, and the heater 28, the fan 29, the temperature controller 31 and the temperature sensor 30 are connected by wires. During the experiment, the target temperature was set to 28°C by the temperature setting plus or minus buttons on the temperature controller 31. The temperature controller 31 will control whether the heater 28 and the fan 29 are working based on the accurate real-time temperature data feedback from the temperature sensor 30 to maintain the stability of the internal ambient temperature of the box body 1.

[0046] In the present invention, the optical imaging system 3 includes an area light source 13 disposed directly below the interior of the box 1. A plurality of cameras 14 are evenly arranged on the surface of the area light source 13 corresponding to the center position of the porous plate 15. The area light source 13 is connected to the interior of the box 1 directly below via a plurality of small telescopic rods, so that the height of the area light source 13 can be adjusted by the small telescopic rods. The luminous area of the area light source 13 is 600mm×600mm (length×width), and the parameters of the camera 14 are: operating temperature: -20℃-70℃, maximum resolution: 4556*3496, operating voltage: 5V, focal length range: 100mm-400mm. During the experiment, the experimenter can monitor the life activity status of the zebrafish larvae at any time through the real-time images transmitted by the camera 14 through the control terminal.

[0047] In the present invention, the entire housing 1 is constructed from a frame structure 7 and acrylic panels. Its overall dimensions are 1080mm × 900mm × 600mm (length × width × height). The housing is constructed from an aluminum alloy frame structure 7 and double-sided black matte acrylic panels. The aluminum alloy frame structure 7 is a rectangular parallelepiped structure and serves as the primary support structure for the entire housing 1, ensuring its structural stability. The black matte acrylic panels are 5mm thick and are manufactured using laser cutting technology. The connections between the frame structure 7 and the housing 1 are secured with screws.

[0048] The present invention also provides a technical solution, a screening method applied to a high-throughput zebrafish drug screening device, comprising the following steps:

[0049] Step 1: Turn on the temperature control system 27 to ensure that the internal temperature of the box 1 is at a constant temperature of 28°C;

[0050] Step 2: Add zebrafish fry to the holes on the surface of the porous plate 15, and place the porous plate 15 with the zebrafish fry in the high-throughput structure unit 4, and wait for 30 minutes to allow the zebrafish fry to adapt to the internal environment of the box 1;

[0051] Step 3: Prepare multiple drugs to be tested, and add the multiple drugs to be tested into the drug test tubes 22 through the drug taking slot 19;

[0052] Step 4: The automated drug delivery system 5 is driven to move by the automated motion control system 2. When the dropping needle provided at the bottom center of the main structure 24 of the automated drug delivery system moves to the position just above the corresponding hole on the surface of the porous plate 15, the drug to be tested is dropped into the hole on the surface of the porous plate 15 by the automated drug delivery system 5.

[0053] Step 5: Capture the movement behavior images of the zebrafish larvae through the optical imaging system 3 and collect the data;

[0054] Step 6: Conduct behavioral analysis on the collected zebrafish larvae movement behavior images.

[0055] Example: Turn on the temperature control system 27 to ensure that the internal temperature of the box 1 is in a constant temperature state of 28°C; during the experiment, the target temperature is set to 28°C through the temperature setting plus or minus buttons on the temperature controller 31. The temperature controller 31 will control whether the heater 28 and the fan 29 are working based on the accurate real-time temperature data feedback from the temperature sensor 30 to maintain the stability of the internal ambient temperature of the box 1.

[0056] Add zebrafish fry to the holes on the surface of the porous plate 15, and place the porous plate 15 with the zebrafish fry in the high-throughput structure unit 4, and wait for 30 minutes to allow the zebrafish fry to adapt to the internal environment of the box 1;

[0057] Prepare multiple drugs to be tested, and add the multiple drugs to be tested into the drug test tube 22 through the drug taking slot 19;

[0058] The automated motion control system 2 drives the automated drug delivery system 5 to move. The longitudinally and transversely arranged motion single shaft 12 cooperates with the stepper motor 9 to drive the entire automated motion control system 2 to move longitudinally and transversely, thereby realizing the longitudinal and transverse movement of the automated drug delivery system housing 21 outside the motion connection structure 11 arranged on the surface of the transversely arranged motion single shaft 12. When the droplet needle set at the bottom center of the automated drug delivery system main structure 24 moves to the corresponding hole on the surface of the porous plate 15, the liquid micropump 233 inside the automated drug delivery system 5 adds the drug to be tested in the drug test tube 22 to the hole on the surface of the porous plate 15 through the droplet needle; during the droplet addition, the liquid micropump 233 can be controlled by the micropump control circuit board 244 to extract different amounts of different test liquids, thereby realizing the detection of different drug concentration gradients. It ensures that the drug content added to the holes on the surface of different porous plates 15 is different, and ultimately realizes the detection of different drug concentration gradients.

[0059] The optical imaging system 3 captures images of the zebrafish larvae's movement behavior and collects data. During the experiment, the experimenter can monitor the zebrafish larvae's life activity status at any time through the control terminal through the real-time images transmitted by the regional light source 13 and the camera 14;

[0060] Behavioral analysis was performed on the collected images of zebrafish larvae's movement behavior.

[0061] During the experiment, except for adding drugs to the drug test tube 22 and taking out the zebrafish fry, the entire box 1 is in a sealed state at other times.

[0062] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description. It is intended that all variations within the meaning and range of equivalents of the claims be embraced herein, and any reference signs in the claims should not be construed as limiting the claims to which they relate.

[0063] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A device for high-throughput zebrafish drug screening, comprising a housing (1), characterized in that: An optical imaging system (3) is provided directly below the interior of the box (1), a plurality of support rods (6) are symmetrically provided on both sides of the optical imaging system (3), an automated motion control system (2) is provided on the top of the support rods (6) on both sides, a high-throughput structural unit (4) is provided at the bottom center of the automated motion control system (2), an automated drug delivery system (5) is connected to the top of the automated motion control system (2), and a temperature control system (27) is also provided inside the box (1); The automated motion control system (2) comprises a mounting seat arranged at the top of the support rods (6) on both sides, a motion single shaft (12) being longitudinally arranged on the top of the mounting seat, two surfaces of the longitudinally arranged motion single shafts (12) being connected to the bottom of the transversely arranged motion single shaft (12) via motion connection structures (11), and the surface of the transversely arranged motion single shaft (12) being connected to an automated drug delivery system (5) via the motion connection structure (11); The high-throughput structural unit (4) includes a high-throughput platform (17), a surface of which is evenly provided with a plurality of porous plates (15), and each porous plate (15) is symmetrically provided with a plate taking slot (18) on both sides; The automated drug delivery system (5) comprises an automated drug delivery system housing (21) arranged on the outside of a motion connection structure (11) on the surface of the horizontally arranged motion single shaft (12); the bottom end of the interior of the automated drug delivery system housing (21) is symmetrically connected to a delivery power device (20) via an automated drug delivery system main structure (24); and a plurality of drug test tubes (22) are connected to the sides of the two delivery power devices (20) that are away from each other.

2. The high-throughput zebrafish drug screening device according to claim 1, characterized in that: One end of the longitudinally arranged and transversely arranged single motion shaft (12) is provided with a stepping motor (9), and each stepping motor (9) is respectively connected to a position positioning switch (10).

3. The high-throughput zebrafish drug screening device according to claim 1, characterized in that: A plurality of connection holes (16) are symmetrically provided on both sides of the high-throughput platform (17), and the connection holes (16) are connected to the surface of the mounting seat at the top of the support rod (6) in conjunction with fastening bolts.

4. The high-throughput zebrafish drug screening device according to claim 1, characterized in that: A drug taking groove (19) is provided at the top of the automatic drug delivery system housing (21) corresponding to the position of the drug test tube (22), and the automatic drug delivery system housing (21) and the laterally arranged motion connection structure (11) are fixedly connected through positioning holes (23) and fastening bolts.

5. The high-throughput zebrafish drug screening device according to claim 1, characterized in that: The delivery power device (20) comprises a power box body connected to the top of the main structure (24) of the automated drug delivery system, a micropump control circuit board (244) is arranged inside the power box body, one end of the micropump control circuit board (244) is connected to a liquid micropump (233), one end of the liquid micropump (233) is provided with a liquid inlet (222) connected to the drug test tube (22) on the adjacent side, and the other end of the liquid micropump (233) is provided with a liquid outlet (211), and both of the two liquid micropumps (233) are connected to the drip needle arranged at the bottom center of the main structure (24) of the automated drug delivery system through the liquid outlet (211) in conjunction with the delivery tube, and the other end of the micropump control circuit board (244) is provided with a delivery power device power interface (25) and a delivery power device control interface (26).

6. The high-throughput zebrafish drug screening device according to claim 1, characterized in that: The temperature control system (27) includes a hot air interface (8) arranged on the rear side of the box (1), a heater (28) is connected to the outside of the hot air interface (8), a fan (29) is also arranged between the hot air interface (8) and the heater (28), and a temperature sensor (30) is also arranged inside the box (1), and the temperature sensor (30) is electrically connected to the temperature controller (31).

7. The device and method for high-throughput zebrafish drug screening according to claim 1, characterized in that: The optical imaging system (3) includes a regional light source (13) arranged directly below the interior of the box (1), and a plurality of cameras (14) are evenly arranged on the surface of the regional light source (13) at positions corresponding to the center of the porous plate (15).

8. The high-throughput zebrafish drug screening device according to claim 1, characterized in that: The box body (1) is made entirely of a frame structure (7) and an acrylic plate.

9. A screening method for a high-throughput zebrafish drug screening device according to claims 1-8, characterized in that: The following steps are involved: Step 1: Turn on the temperature control system (27) to ensure that the internal temperature of the box (1) is at a constant temperature of 28°C; Step 2: Add zebrafish fry to the holes on the surface of the porous plate (15), and place the porous plate (15) with the zebrafish fry in the high-throughput structure unit (4), and wait for 30 minutes to allow the zebrafish fry to adapt to the internal environment of the box 1; Step 3: preparing a plurality of drugs to be tested, and adding the plurality of drugs to be tested into the drug test tubes (22) through the drug taking slot (19); Step 4: The automated drug delivery system (5) is driven to move by the automated motion control system (2). When the dripping needle provided at the bottom center of the main structure (24) of the automated drug delivery system moves to the position just above the corresponding hole on the surface of the porous plate (15), the drug to be tested is dripped into the hole on the surface of the porous plate (15) by the automated drug delivery system (5); Step 5: Collecting the movement behavior images of the zebrafish larvae through the optical imaging system (3) and collecting the data; Step 6: Conduct behavioral analysis on the collected zebrafish larvae movement behavior images.

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