Zebra fish two-dimensional behavior analysis device and method

Abstract

The invention discloses a zebra fish two-dimensional behavior analysis device and method.The device is characterized in that an object placing platform is arranged at the bottom in a box body, a temperature control small chamber is placed on the object placing platform, a vibration generator and a perforated plate container are arranged in the temperature control small chamber, and the vibration generator is arranged on the side edge in the temperature control small chamber; the perforated plate container is arranged at the bottom in the temperature control chamber, the full-spectrum simulation light source is fixed to the inner side wall of the box body, the infrared LED light source is further arranged at the bottom of the temperature control chamber, the controllable light source is arranged between the bottom of the temperature control chamber and the storage platform, the shooting supporting platform is arranged above the storage platform, and the fan device is arranged on the outer side of the box body. A circuit control board is further arranged on the outer side of the box body. Long-term continuous simulation is achieved through the temperature control chamber, the full-spectrum simulation light source, the infrared LED light source and the controllable light source, the experiment efficiency is improved, equipment expansion can be achieved, and diversified experiment requirements are met.

Description

Technical Field

[0001] The present invention relates to the technical field of drug screening, and in particular to a device and method for analyzing two-dimensional behavior of zebrafish. Background Art

[0002] Zebrafish are biologically very similar to humans, sharing most of their genes. 70% of protein-coding human genes are related to genes found in zebrafish, and 84% of genes known to be associated with human diseases have zebrafish counterparts. Furthermore, zebrafish spawn year-round, are extremely low-cost to rear, and have minimal experimental ethical pressures, allowing for high-throughput screening. Therefore, investigating the behavioral responses of zebrafish to different drug stimulation conditions is of great significance for the study of human diseases.

[0003] Existing technologies rely on fewer variables to regulate zebrafish behavior and conduct drug screening. For example, CN114680069A, published on July 1, 2022, is titled "A Zebrafish Behavior and Movement Experimental Device." It uses a zebrafish behavior and movement experimental device in which a water pump adjusts the water flow rate through a speed regulator to provide a stable water flow, making it convenient to control variables to study the behavior and movement of zebrafish. Several experimental tanks are provided to adjust different experimental variables to study the effects of water flow rate and food intake on zebrafish movement disorders. By controlling variables such as water flow rate and zebrafish food feeding, the behavior and movement of zebrafish are detected. This type of method has a single detection function and cannot simultaneously capture and analyze movement trajectories.

[0004] Therefore, existing technologies lack the ability to simultaneously monitor multiple zebrafish under different environmental (drug) conditions and complete high-throughput experiments. Various chambers must be set up to study the behavior and movement trajectories of zebrafish under different experimental variables. There is no complete equipment that can simultaneously maintain a suitable growth environment for zebrafish over a long period of time, control different experimental variables to study the movement trajectories of zebrafish under certain stimuli, and analyze and process the results. When there are too few controllable factors in an experiment, errors are likely to occur. Summary of the Invention

[0005] The present invention provides a two-dimensional zebrafish behavior analysis device and method to address the existing challenges of simultaneously monitoring multiple zebrafish under different environmental conditions and completing high-throughput experiments. This requires setting up various chambers to study the behavioral motion trajectories of zebrafish under different experimental variables. No single comprehensive device can simultaneously maintain a suitable growth environment for zebrafish over a long period of time, control different experimental variables to study the motion trajectories of zebrafish under specific stimuli, and simultaneously analyze and process the results. When there are too few controllable factors in an experiment, errors are easily generated.

[0006] In one aspect, an embodiment of the present invention provides a zebrafish two-dimensional behavior analysis device, comprising:

[0007] A box body is provided with a storage platform at the bottom of the box body, a temperature-controlled chamber is placed on the storage platform, a vibration generator and a multi-hole plate container are provided in the temperature-controlled chamber, the vibration generator is provided on the inner side of the temperature-controlled chamber, the multi-hole plate container is provided at the bottom of the temperature-controlled chamber, the full-spectrum simulation light source is fixed on the inner wall of the box body, an infrared LED light source is also provided at the bottom of the temperature-controlled chamber, a controllable light source is provided between the bottom of the temperature-controlled chamber and the storage platform, a shooting support platform is provided above the storage platform, the shooting support platform is used to support the shooting device, a fan device is provided on the outside of the box body, and a circuit control board is also provided on the outside of the box body, and the circuit control board signal is connected to the vibration generator, the full-spectrum simulation light source, the infrared LED light source, the controllable light source, the shooting device, the fan device and the host computer.

[0008] In a possible implementation, the circuit control board is connected to a host computer via a USB connector signal provided on the outside of the box.

[0009] In a possible implementation, the storage platform is a hollow platform, and the infrared LED light source and the controllable light source are arranged between the storage platform and the box to illuminate the temperature-controlled chamber.

[0010] In a possible implementation, the full-spectrum simulation light source is provided with four adjustable LED light sources: white light, UV light, blue light, and dark blue light.

[0011] In a possible implementation, the temperature control chamber is further provided with a temperature and humidity control unit, a water storage tank, a vent and a control channel.

[0012] In a possible implementation, a temperature and humidity sensor is provided in the temperature and humidity control unit, and the temperature and humidity control unit is connected to the circuit control board via the control channel signal.

[0013] On the other hand, an embodiment of the present invention provides a method for analyzing two-dimensional behavior of zebrafish, comprising:

[0014] The temperature and humidity of the temperature-controlled chamber are adjusted to the living temperature and humidity of zebrafish through the temperature and humidity control unit and the fan device;

[0015] After the zebrafish is placed in a multi-well plate container, the multi-well plate container is placed in a temperature-controlled chamber;

[0016] Use the circuit control board to connect an external power supply to power the controllable light source and shooting device and turn them on;

[0017] The host computer controls the shooting device to collect images to obtain zebrafish movement data;

[0018] The zebrafish movement data is analyzed by a host computer to obtain the zebrafish movement trajectory.

[0019] In a possible implementation, analyzing the zebrafish motion data by a host computer to obtain the zebrafish motion trajectory is performed by marking the zebrafish head center of mass as a tracking point for analysis to obtain the zebrafish motion trajectory.

[0020] The device and method for analyzing two-dimensional zebrafish behavior in the present invention have the following advantages:

[0021] (1) Long-term continuous simulation is achieved through a temperature-controlled chamber, a full-spectrum simulated light source, an infrared LED light source, and a controllable light source.

[0022] (2) The measurement equipment and variable control equipment are set together in a temperature-controlled chamber, which improves the efficiency of the experiment.

[0023] (3) By increasing the controllability of variables through full-spectrum simulated light sources, infrared LED light sources and controllable light sources, the equipment can be expanded to meet diverse experimental needs. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0025] Figure 1 A schematic diagram of a zebrafish two-dimensional behavior analysis device provided by an embodiment of the present invention;

[0026] Figure 2 A schematic cross-sectional view of a device for analyzing two-dimensional zebrafish behavior provided by an embodiment of the present invention;

[0027] Figure 3 A schematic diagram of the temperature control chamber structure of a zebrafish two-dimensional behavior analysis device provided by an embodiment of the present invention;

[0028] Figure 4 A flowchart of a zebrafish two-dimensional behavior analysis method provided by an embodiment of the present invention;

[0029] Figure 5 An image diagram of the multi-well plate alignment of a zebrafish two-dimensional behavior analysis device provided by an embodiment of the present invention;

[0030] Figure 6 This is a convenient analysis image of a multi-well plate of a zebrafish two-dimensional behavior analysis device provided by an embodiment of the present invention;

[0031] Figure 7 A schematic diagram of a two-dimensional behavior analysis device for zebrafish provided in an embodiment of the present invention; and a diagram of zebrafish trajectories in a multi-well plate.

[0032] Figure 8 A quantified graph of zebrafish movement distance in a zebrafish two-dimensional behavior analysis device provided in an embodiment of the present invention.

[0033] Explanation of the numbers in the figure: 1. Box; 2. Storage platform; 3. Temperature control chamber; 31. Temperature and humidity control unit; 32. Water tank; 33. Vent; 34. Control channel; 4. Vibration generator; 5. Multi-hole plate container; 6. Full-spectrum simulation light source; 7. Infrared LED light source; 8. Controllable light source; 9. Circuit control board; 10. Camera device; 11. Fan device; 12. USB connector; 14. Camera support platform; 15. Host computer. DETAILED DESCRIPTION

[0034] 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.

[0035] Figure 1 Schematic diagram of a two-dimensional zebrafish behavior analysis device provided in an embodiment of the present invention. An embodiment of the present invention provides a two-dimensional zebrafish behavior analysis device, comprising:

[0036] The box body 1 is provided with a storage platform 2 at the bottom of the box body 1, and a temperature-controlled chamber 3 is placed on the storage platform 2. A vibration generator 4 and a porous plate container 5 are provided in the temperature-controlled chamber 3. The vibration generator 4 is arranged on the inner side of the temperature-controlled chamber 3, and the porous plate container 5 is arranged at the bottom of the temperature-controlled chamber 3. The full-spectrum simulation light source 6 is fixed to the inner wall of the box body 1, and an infrared LED light source 7 is also provided at the bottom of the temperature-controlled chamber 3. A controllable light source 8 is provided between the bottom of the temperature-controlled chamber 3 and the storage platform 2. A shooting support platform 14 is provided above the storage platform 2, and the shooting support platform 14 is used to support the shooting device 10. A fan device 11 is provided on the outside of the box body 1, and a circuit control board 9 is also provided on the outside of the box body 1. The circuit control board 9 is signal-connected to the vibration generator 4, the full-spectrum simulation light source 6, the infrared LED light source 7, the controllable light source 8, the shooting device 10, the fan device 11 and the host computer 15.

[0037] The circuit control board 9 is connected to the host computer 15 via a USB connector 12 arranged on the outside of the box 1 .

[0038] The storage platform 2 is a hollow platform, and the infrared LED light source 7 and the controllable light source 8 are arranged between the storage platform 2 and the box body 1 to illuminate the temperature control chamber 3.

[0039] The full-spectrum simulation light source 6 is provided with four adjustable LED light sources of white light, UV light, blue light and dark blue light.

[0040] The temperature control chamber 3 is further provided with a temperature and humidity control unit 31 , a water storage tank 32 , a vent 33 and a control channel 34 .

[0041] The temperature and humidity control unit 31 is provided with a temperature and humidity sensor, and the temperature and humidity control unit 31 is connected to the circuit control board 9 via the control channel 34 signal.

[0042] For example, Figure 1 、 2 As shown in Figure 3, the size of the box 1 is set to multiple specifications. The size of the box 1 and the size of the temperature-controlled chamber are set according to the number of holes in the multi-well plate container 5. The box 1 is made of black polylactic acid material to prevent the influence of external light. The shooting device 10, the multi-well plate container 5 and the controllable light source 8 are vertically distributed. The temperature-controlled chamber 3, the shooting system 10, the controllable light source 8, the circuit control board 9 and the fan device 11 are placed in the same box 1 to reduce the impact of the outside world on the zebrafish culture environment. The device has a high degree of integration, is easy to operate, and reduces experimental errors. The storage platform 2 is a hollow platform composed of four cylindrical metal supports and a layer of aluminum alloy platform, which provides stable support for the temperature-controlled chamber 3 above. The temperature-controlled chamber 3 is located in the middle part of the box. A porous plate container 5 is provided in the middle part of the temperature-controlled chamber 3, and a vibration generator 4 is provided on the side of the porous plate container 5. The vibration generator 4 is controlled by the circuit control board 9 to generate vibration at a certain frequency in the temperature-controlled chamber 3, thereby stimulating the zebrafish in the porous plate container 5 to keep moving.

[0043] The shape of the multi-well plate is rectangular, and the shape of the holes is circular. The size and number of holes of the multi-well plate are set in multiple specifications, and multi-well plates such as 96-well plates and 24-well plates can be used.

[0044] The temperature and humidity control unit 31 is connected to and powered by an external power source. A temperature and humidity sensor installed within the temperature and humidity control unit 31 detects the temperature and humidity within the temperature-controlled chamber 3. The circuit control board 9 then controls the fan device 11 to achieve controllable temperature and humidity within the temperature-controlled chamber 3. The circuit control board 9 controls the fan device 11 to activate in a dark environment, providing fill light for the camera 10, allowing it to capture and record experimental data even in the dark.

[0045] A full-spectrum simulated light source 6 is located on the side wall of the housing 1 and simulates the process from sunrise to sunset using an adjustable LED light source (white light, UV violet light, blue light, and dark blue light). An infrared LED light source 7 is located at the bottom of the housing 1 and contains four 850nm infrared LED lamp beads, which provide a field of view for the camera 10 in dark environments. A fan device 11 is embedded in the rear wall of the housing 1. The fan device 11 is connected to an external temperature and humidity control box and is powered by an external power supply.

[0046] In one possible embodiment, the experimental sample is a juvenile zebrafish, and a single camera, a single multi-well plate and a single surface light source are used; the multi-well plate used is a 24-well plate, and a total of 24 zebrafish are cultured and observed. The light source used is a surface light source, with an operating voltage of 1.8-3.6V, an operating current of 1.5-20mA, and a luminous color of white light. The temperature is adjusted to 28°C and the humidity is 99%. The shooting device 10 is a CCD camera. After the shooting device 10 obtains the picture, it is transmitted to the host computer 15 for analysis through the circuit control board 9. During this period, the image is as follows: Figure 5 、 6 , 7, and 8.

[0047] Figure 4 Flowchart of a two-dimensional zebrafish behavior analysis device provided by an embodiment of the present invention; an embodiment of the present invention provides a two-dimensional zebrafish behavior analysis device, comprising:

[0048] The temperature and humidity of the temperature-controlled chamber 3 are adjusted to the living temperature and humidity of zebrafish by the temperature and humidity control unit 31 and the fan device 11;

[0049] After the zebrafish is placed in the multi-well plate container 5, the multi-well plate container 5 is placed in the temperature-controlled chamber 3;

[0050] Use the circuit control board 9 to connect to an external power source to power the controllable light source 8 and the shooting device 10 and turn them on;

[0051] The host computer 15 controls the shooting device 10 to collect images to obtain zebrafish movement data;

[0052] The host computer 15 analyzes the zebrafish motion data to obtain the zebrafish motion trajectory.

[0053] In a possible embodiment, the zebrafish motion trajectory obtained by analyzing the zebrafish motion data by the host computer 15 is obtained by marking the zebrafish head center of mass as a tracking point for analysis.

[0054] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including all changes and modifications that fall within the scope of the present invention and the preferred embodiments.

[0055] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A zebrafish two-dimensional behavior analysis device, characterized in that: include: A box (1) is provided with a storage platform (2) at the bottom of the box (1), a temperature-controlled chamber (3) is placed on the storage platform (2), a vibration generator (4) and a multi-hole plate container (5) are provided in the temperature-controlled chamber (3), the vibration generator (4) is provided on the inner side of the temperature-controlled chamber (3), the multi-hole plate container (5) is provided at the bottom of the temperature-controlled chamber (3), a full-spectrum simulation light source (6) is fixed to the inner side wall of the box (1), an infrared LED light source (7) is further provided at the bottom of the temperature-controlled chamber (3), and the bottom of the temperature-controlled chamber (3) is connected to the A controllable light source (8) is provided between the storage platforms (2), a shooting support platform (14) is provided above the storage platforms (2), and the shooting support platform (14) is used to support the shooting device (10). A fan device (11) is provided on the outside of the box (1), and a circuit control board (9) is also provided on the outside of the box (1). The circuit control board (9) is connected to the vibration generator (4), the full-spectrum simulation light source (6), the infrared LED light source (7), the controllable light source (8), the shooting device (10), the fan device (11) and the host computer (15) through signals.

2. A zebrafish two-dimensional behavior analysis device according to claim 1, characterized in that: The circuit control board (9) is connected to the host computer (15) via a USB connector (12) arranged outside the box (1).

3. The zebrafish two-dimensional behavior analysis device according to claim 1, characterized in that: The storage platform (2) is a hollow platform, and the infrared LED light source (7) and the controllable light source (8) are arranged between the storage platform (2) and the box (1) to illuminate the temperature-controlled chamber (3).

4. The device for analyzing zebrafish two-dimensional behavior according to claim 1, wherein: The full-spectrum simulation light source (6) is provided with four adjustable LED light sources: white light, UV light, blue light, and dark blue light.

5. The zebrafish two-dimensional behavior analysis device according to claim 1, characterized in that: The temperature control chamber (3) is further provided with a temperature and humidity control unit (31), a water storage tank (32), a vent (33) and a control channel (34).

6. The device for analyzing zebrafish two-dimensional behavior according to claim 5, characterized in that: A temperature and humidity sensor is provided in the temperature and humidity control unit (31), and the temperature and humidity control unit (31) is connected to the circuit control board (9) via the control channel (34) signal.

7. A zebrafish two-dimensional behavior analysis method, characterized in that: include: The temperature and humidity of the temperature-controlled chamber (3) are adjusted to a temperature and humidity suitable for zebrafish survival by using a temperature and humidity control unit (31) and a fan device (11); After placing the zebrafish into the multi-well plate container (5), the multi-well plate container (5) is placed into a temperature-controlled chamber (3); Use the circuit control board (9) to connect an external power source to power the controllable light source (8) and the shooting device (10) and turn them on; The shooting device (10) is controlled by a host computer (15) to acquire images and obtain zebrafish movement data; The zebrafish movement data is analyzed by a host computer (15) to obtain the zebrafish movement trajectory.

8. A zebrafish two-dimensional behavior analysis method according to claim 7, characterized in that: The zebrafish motion data is analyzed by the host computer (15) to obtain the zebrafish motion trajectory, and the zebrafish motion trajectory is obtained by marking the zebrafish head center of mass as a tracking point for analysis.

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