Device for cognitive analysis of social behavior of zebrafish larvae

Abstract

The utility model discloses a zebra fish juvenile fish social cognition behavior analysis device, and the device comprises an analyzer body, and the body is mainly composed of a box, a flue mechanism, a tray, an illumination mechanism and a camera. Its core lies in, the flue mechanism adopts detachable design, and a plurality of independent flues that can be flexibly separated by movable partition plates are arranged on the flue mechanism, parallel experiment capacity of high flux and multiple scenes is realized, and opaque side plates are arranged between adjacent flues. The illumination mechanism is arranged below the tray, has a plurality of illumination areas that can be independently programmed and controlled, can provide light stimulation of multiple colors and modes, and the camera is arranged directly above the flue, is used for collecting juvenile fish behavior images and transmitting to the computer for automatic analysis. The utility model integrates environmental control and behavior observation, solves the problems of low flux, rough environmental regulation and strong analysis subjectivity of the prior art, and significantly improves the efficiency and accuracy of zebra fish juvenile fish social behavior research.

Description

Technical Field

[0001] This utility model relates to the field of life science research technology, specifically to a device for analyzing the social behavior and cognition of zebrafish juveniles. Background Technology

[0002] Zebrafish, as an important model organism, are widely used in developmental biology, neuroscience, and drug screening due to their high genetic homology with humans, transparent embryonic development, and short reproductive cycle. In particular, the juvenile zebrafish, with their neuroplasticity and rich social behavioral patterns, provide an ideal model for studying social behavioral development and the pathogenesis of neuropsychiatric disorders.

[0003] However, current experimental setups and techniques for studying the social behavior of zebrafish juveniles still have significant limitations, severely restricting the efficiency and depth of research. These limitations are mainly reflected in the following aspects: First, the experimental throughput and efficiency are low. Traditional research often uses a single aquarium or simple partitioning device, which can only conduct single-group or a small number of control experiments at a time. This method is not only time-consuming and labor-intensive, but also makes it difficult to ensure the consistency of experimental conditions between different batches, resulting in poor data comparability and failing to meet the needs of modern life sciences for high-throughput and standardized data collection.

[0004] Second, the experimental environment is poorly controlled, lacking standardization and flexibility. The behavior of juvenile zebrafish is easily affected by environmental factors such as light and temperature. Existing equipment often lacks a sophisticated light control system, making it impossible to simulate complex and changing light environments or provide stable and uniform experimental conditions. In addition, the fixed layout of the experimental space makes it difficult to flexibly reconstruct experimental scenarios according to different experimental objectives (such as social preferences, cognitive decision-making, aggressive behavior, etc.), thus limiting the diversity of research paradigms.

[0005] Third, the automation and precision of behavioral observation and data analysis are insufficient. Many studies still rely on manual observation and recording, which is highly subjective and cannot capture and quantify the rapid and subtle behavioral dynamics of juvenile fish (such as instantaneous approaching, avoidance, or posture adjustments). Although some commercial behavioral analysis systems have video recording capabilities, their algorithms are usually designed for tracking the trajectory of a single fish in open water, making it difficult to accurately identify and analyze the details of social interactions among multiple juvenile fish in complex channel environments with physical separation.

[0006] Therefore, there is an urgent need in this field for an integrated solution that can enable high-throughput, parallel experiments on the social behavior of zebrafish juveniles, provide precise, controllable, and flexible environmental stimuli, and possess automatic and accurate image capture and behavioral analysis capabilities, in order to overcome the shortcomings of existing technologies and promote the in-depth development of related research. Utility Model Content

[0007] In view of the above-mentioned defects or deficiencies in the existing technology, it is desirable to provide a zebrafish juvenile social behavior cognitive analysis device to solve the technical problems that existing zebrafish juvenile social behavior cognitive analysis devices cannot automatically observe and analyze zebrafish juveniles, and cannot accurately and efficiently analyze the social behavior of zebrafish juveniles under different lighting conditions.

[0008] According to the technical solution provided in the embodiments of this application, a zebrafish juvenile social cognitive behavior analysis device includes an analyzer body. The analyzer body includes a housing, a passageway mechanism, a tray, a lighting mechanism, and a camera; The passageway mechanism includes a passageway plate, which is detachably mounted on the top of the tray, and multiple independent passageway channels are arranged side by side on the passageway plate; The lighting mechanism is located below the tray and is used to provide controllable lighting to the passageway. The camera is installed at the top of the inside of the enclosure and directly above the passageway panel, and is used to film the behavior of juvenile zebrafish in the passageway.

[0009] In this invention, there are ten passageways, and each passageway is a rectangular groove structure.

[0010] In this invention, at least one movable partition is provided in the passageway, which divides the passageway into multiple experimental sections.

[0011] In this invention, the movable partition is installed in the slot of the passageway plate by snap-fit ​​or plug-in connection.

[0012] In this invention, a side panel is provided between two adjacent passageways. The side panel is made of an opaque material to achieve visual isolation.

[0013] In this invention, the lighting mechanism includes a lighting plate, on which multiple independent lighting areas are provided, and each lighting area can emit at least two different colors of light.

[0014] In this invention, there are three lighting areas arranged side by side along the transverse direction of the lighting panel; each lighting area can independently emit white, green, blue or red light.

[0015] In this invention, the front end of the box is provided with an opening, and a door that can be raised and lowered in the vertical direction is installed at the opening.

[0016] In this invention, the camera is connected to an external computer via a data cable, and the external computer is equipped with analysis software for analyzing zebrafish behavior images.

[0017] In this invention, the bottom of the tray has a conical structure. In summary, the beneficial effects of this application are as follows: 1. By setting a tunnel mechanism inside the analyzer's housing, the tunnel plate on the mechanism is placed on a tray via fixing pins. Ten rectangular tunnel channels are set on the tunnel plate, allowing the analyzer to conduct different social behavior experiments on zebrafish larvae simultaneously. This tunnel design allows for flexible adjustment of the tunnel's spatial layout according to different experimental needs during the experiment. For example, different experimental items or different experimental conditions can be set in different parts. For example, a mirror can be added to the lower part for social experiments to observe whether the larvae's social tendencies are playing with themselves, attacking their reflection in the mirror, or interacting with other larvae. Second, the side panels of the tunnels are made opaque. This feature ensures that the different tunnels cannot see each other, which can effectively prevent the fish in the next tunnel from interfering with the experiment. It provides an independent and undisturbed experimental environment for the juvenile fish in each tunnel, which is conducive to the accurate observation and analysis of the behavior of the juvenile fish and the juvenile fish group. Third, by setting up an illumination mechanism at the bottom of the analyzer housing, and keeping the illumination mechanism a certain distance from the passage plate, the heat generated by the illumination mechanism will not affect the temperature during the observation process. At the same time, three illumination areas are set up on the illumination mechanism, so that the three illumination areas can emit four colors of light: white, green, blue and red. This allows for diverse lighting changes within the analyzer to elicit different behavioral behaviors in juvenile fish, enabling a more comprehensive study of the behavioral patterns of juvenile fish. Attached Figure Description

[0018] Other features, objects, and advantages of this application will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings: Figure 1 This is a front-view three-dimensional structural diagram of the present invention; Figure 2 This is a rear-view three-dimensional structural diagram of the present invention; Figure 3 This is a schematic diagram of the cross-sectional structure of the box body of this utility model; Figure 4 This is a schematic diagram of the three-dimensional structure of the tunnel of this utility model; Figure 5 This is a top view of the lighting mechanism of this utility model.

[0019] The following components are labeled in the diagram: Analyzer body 100, Cabinet 110, Passageway mechanism 120, Passageway plate 121, Passageway 122, Movable partition 123, Side plate 124, Tray 130, Lighting mechanism 140, Lighting panel 141, Lighting area 142, Camera 150, Connection port 151, Door 160, Computer 170. Detailed Implementation

[0020] The present application will now be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the relevant utility model and not intended to limit the scope of the utility model. Furthermore, it should be noted that, for ease of description, only the parts relevant to the utility model are shown in the accompanying drawings.

[0021] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. This application will now be described in detail with reference to the accompanying drawings and embodiments.

[0022] The zebrafish juvenile social behavior cognitive analysis device includes an analyzer body 100; the analyzer body 100 includes a housing 110, a passage mechanism 120, a tray 130, a lighting mechanism 140, and a camera 150; the housing 110 has a rectangular structure with an opening at its front end, and a door 160 that can move up and down vertically is installed at the opening. In the illustration of this scheme, the door 160 is opened by a conventional pull-out method (in subsequent improvements, the top of the door 160 can be connected to a piston cylinder via a linkage rod, so that the linkage rod extends along the gas input of the piston cylinder, thereby opening the door 160 along the opening, which is convenient for electrified operation), which facilitates the replacement of the passage mechanism 120, and also facilitates the cleaning and maintenance of the passage channel 122 during the experiment to meet different experimental needs.

[0023] like Figure 3 and Figure 4As shown, the passageway mechanism 120 includes a passageway panel 121. The rectangular passageway panel 121 is detachably mounted on the top of the tray 130 via fixing pins. Ten passageway channels 122 are provided on the surface of the passageway panel 121, arranged side-by-side and spaced apart along the passageway panel 121. Each passageway channel 122 is a rectangular groove with a length of 50mm, a width of 5mm, and a height of 5mm. Each passageway channel 122 is movably divided into an upper, middle, and lower section by two partitions 123, which are movably engaged in slots. A side panel 124, made of opaque material, is provided between two adjacent passageway channels 122. This design of the passageway channels 122 makes the passageway... During the experiment, the spatial layout of the passageway 122 can be flexibly adjusted according to different experimental needs. For example, different experimental items or different experimental conditions can be set in different parts (for social experiments, a mirror can be added to the lower part to observe whether the social tendencies of the juvenile fish are to play with themselves, attack their reflection in the mirror, or interact with other juvenile fish). The side panels 124 of the passageway are opaque, which ensures that the various passageways 122 cannot see each other, effectively preventing the zebrafish juveniles in adjacent passageways 122 from interfering with the experiment. This provides an independent and undisturbed experimental environment for the zebrafish juveniles in each passageway 122, which is conducive to the accurate observation and analysis of the social behavior of the juvenile fish and the juvenile fish group.

[0024] like Figure 3 and Figure 5 As shown, the lighting mechanism 140 is installed at intervals on the bottom of the tray 130 on the housing 110. The bottom of the tray 130 has a conical cavity structure. The lighting mechanism 140 includes a lighting plate 141, which is installed at a certain distance from the passageway plate 121. The lighting plate 141 has three lighting areas 142, which are arranged side by side along the horizontal direction of the lighting plate 141. The light source size of the lighting plate 141 is adapted to the passageway plate 121, with a length of 55 mm and a width of 50 mm. The light in each horizontal row of lighting areas 142 can be adjusted to four colors: white, green, blue, and red through a program. This light source design... It can provide precise lighting conditions for different types of experiments. In social experiments, the light is adjusted to be completely white to provide a relatively uniform and stable lighting environment for the social behavior of juvenile fish. In cognitive experiments, the light sources in the upper and lower lighting areas 142 are adjusted to blue and red to stimulate the juvenile fish and observe their cognitive responses using the characteristics of different colored light. In behavioral experiments, the upper and lower lighting areas 142 are adjusted to white light, while the light color in the middle lighting area 142 is constantly changing between white, green, blue, and red. This diverse lighting variation can induce different behavioral behaviors in juvenile fish, allowing for a more comprehensive study of their behavioral patterns.

[0025] like Figure 2 and Figure 3As shown, camera 150 is fixedly installed inside the top of the enclosure 110, directly above the passageway plate 121. Camera 150 uses high resolution and high frame rate, enabling it to clearly capture the extremely subtle and complex behaviors of zebrafish juveniles, such as rapid interactive behaviors and subtle posture changes. A connection port 151 is provided on the top of camera 150, which is electrically connected to computer 170 via a connection cable. Camera 150 is also externally connected to computer 170 via a data cable, transmitting the captured image data to computer 170 in real time. At the same time, computer 170 is equipped with dedicated image analysis software. This software has been optimized for the multi-passage structure of this application, and can effectively process and analyze image data obtained from different passageway channels 122, accurately identify and track the behavioral trajectory and movement status of each juvenile fish in the passageway, providing strong support for subsequent experimental data statistics and analysis.

[0026] The working principle of this zebrafish juvenile social behavior cognitive analysis device is as follows: During the analysis and observation of social behavior in zebrafish juveniles using the analyzer, a passageway mechanism 120 is installed within the analyzer's housing 110. The passageway plate 121 of the mechanism 120 is placed on a tray 130 via fixing pins. Ten rectangular passageways 122 are arranged on the passageway plate 121, and each passageway 122 is equipped with a removable partition 123. This allows the analyzer to flexibly adjust the spatial layout of the passageways according to different experimental needs. The side plate 124 between adjacent passageways 122 is configured as... The tunnel is opaque to prevent adjacent passageways 122 from seeing each other, providing an independent and undisturbed experimental environment for the juvenile fish in each passageway. This facilitates accurate observation and analysis of the behavior of the juvenile fish and their groups. An illumination mechanism 140 is installed at the bottom of the analyzer housing 110, with the illumination plate 141 on the mechanism 140 spaced a certain distance from the passageway plate 121. This ensures that the heat generated by the illumination mechanism 140 will not affect the temperature during observation. Three illumination areas 142 are provided on the illumination plate 141 to allow for illumination in all three areas. The system emits four colors of light: white, green, blue, and red. During socialization experiments, the light is set to pure white to provide a relatively uniform and stable lighting environment for the juvenile fish's social behavior. In cognitive experiments, the upper and lower lighting areas (142) are set to blue and red light to stimulate the juvenile fish and observe their cognitive responses. During behavioral experiments, the upper and lower lighting areas (142) are set to white light, while the central lighting area (142) continuously changes between white, green, blue, and red light colors to evoke different responses in the juvenile fish through diverse lighting variations. Behavioral performance is observed to more comprehensively study the behavioral patterns of juvenile fish. Meanwhile, a camera 150 is installed at the top center of the tank 110. The camera 150 uses high resolution and high frame rate equipment, which can clearly capture the extremely subtle and complex and variable behaviors of zebrafish juveniles in each passageway 122, and transmit the acquired data to the electrically connected computer 170 in a timely manner. The computer 170, with the help of specialized image analysis software, can transmit and process image data in real time, accurately identify and track the behavioral trajectory of juvenile fish, and improve data processing efficiency and analysis accuracy.

[0027] 1. Regarding the box body 110 and the door body 160 The enclosure 110 is a rectangular structure made of opaque, heat-insulating material (such as ABS engineering plastic or acrylic), with external dimensions of approximately 400mm long × 300mm wide × 300mm high, used to isolate it from external environmental and light interference. A rectangular opening is provided at the front of the enclosure, and its door 160 is preferably driven by a set of silent linear guide rails and a stepper motor to achieve smooth vertical lifting. In another embodiment, a manual lifting structure can also be used. This design facilitates quick replacement of the passageway mechanism, feeding, or cleaning and maintenance by experimental personnel, while minimizing disturbance to the stable environment inside the enclosure.

[0028] 2. Regarding the passageway mechanism 120 and pallet 130 The tunnel mechanism 120 is the key to achieving high-throughput parallel experiments in this device. The tunnel plate 121 is made of opaque, biocompatible materials such as black PVC or Delrin (polyoxymethylene), and is precision-machined. It achieves rapid and accurate positioning and detachable installation by cooperating with the fixing pins on the tray 130 through the positioning pin holes at the four corners.

[0029] The passageway panel 121 is precisely machined with ten identical passageways 122. Each passageway 122 is a rectangular waterway with an internal cavity size of 50mm long × 5mm wide × 5mm high. This size is optimized to accommodate multiple juvenile fish for social interaction while limiting their free swimming range, making it easier for the camera to capture effective behavior.

[0030] To achieve flexibility in experimental paradigms, each passageway 122 is divided into three experimental sections—upper, middle, and lower—along its length by two movable partitions 123. The movable partitions 123 are made of transparent or translucent material (such as acrylic) and have protrusions at their bottom for secure insertion into pre-set slots on the inner side of the passageway panel 121. Researchers can insert or remove partitions as needed to create connected or separated experimental spaces. For example, when studying mirror-image stimulus responses, a mirror can be inserted into the lower section; when studying social preferences, different types of stimuli can be placed in different sections using partitions.

[0031] Most importantly, adjacent passageways 122 are completely separated by opaque side panels 124. This design ensures that during the experiment, zebrafish juveniles in either passageway cannot see the situation in their adjacent passageway, thus effectively avoiding visual interference and guaranteeing the independence and accuracy of each set of experimental data.

[0032] The bottom of the tray 130 is designed with a conical cavity structure with an inclination angle greater than 5 degrees. This structure facilitates the rapid collection of water and impurities to the lowest point for discharge during drainage or cleaning, ensuring that the interior of the device is easy to clean and dry, and maintaining experimental hygiene.

[0033] 3. Regarding the lighting facility 140 The lighting mechanism 140 is fixedly installed below the tray 130, maintaining a distance of approximately 10-15 mm from the bottom surface of the passageway plate 121. This distance ensures that the light evenly covers all passageways, and also utilizes air convection to prevent the heat generated by the lighting from being directly conducted to the experimental water, thereby maintaining a stable water temperature.

[0034] The core of the lighting mechanism is a custom-designed high-power LED lighting board 141 with a light-emitting surface size of 55mm × 50mm, adapted to the projected area of ​​the passageway panel 121. This lighting board 141 is divided into three independent lighting zones 142 in terms of hardware and control logic. Each lighting zone 142 can be independently programmed via an external control system such as an Arduino or STM32 microcontroller, precisely controlling its on / off state, brightness, and color temperature, emitting white, green, blue, and red light.

[0035] Application examples of lighting control: Social interaction experiment: The upper, middle and lower lighting areas 142 were uniformly set to stable white cool light to provide a standard, unbiased lighting environment for all passageways, focusing on observing the natural interactions between juvenile fish.

[0036] Cognitive Experiment: The upper and lower lighting areas 142 can be set to continuous blue light, while the middle area can be set to pulsed red light. The zebrafish's innate preference for or stress response to different colors and frequencies of light can be used to test their learning, memory, and decision-making abilities.

[0037] Behavioral preference experiment: The upper and lower areas were set to white background light, while the light color of the middle area was cyclically switched between four colors: white, green, blue, and red according to a preset program. Through dynamic light environment stimulation, the behavioral preferences and adaptive changes of juvenile fish were induced and observed.

[0038] 4. Regarding image acquisition and analysis systems The camera 150 is fixed to the center of the top of the inside of the housing 110 by a universal bracket, ensuring that its lens faces and covers the entire passageway 121. The camera 150 preferably has an industrial-grade CMOS camera with a resolution of 1080P (or higher) and a frame rate of 60 frames per second or higher, so as to ensure that it can clearly capture the fast and subtle swimming postures and social behaviors (such as tail wagging, attacking, approaching, etc.) of zebrafish juveniles.

[0039] The connection port 151 on the top of the camera 150 is connected to an external computer 170 (or embedded processing unit) via a high-quality shielded data cable. The computer 170 runs dedicated zebrafish behavior analysis software. This software integrates background subtraction, multi-target tracking, and behavior recognition algorithms, and is capable of: Real-time tracking: Simultaneously track the center-of-gravity movement trajectory of multiple juvenile fish in ten channels.

[0040] Parameter extraction: Automatic quantification calculation includes key behavioral parameters such as swimming speed, activity distance, dwell time in each section, number and duration of social contact.

[0041] Data output: The processed data is output in the form of charts or tables for researchers to conduct statistical analysis, which greatly improves the efficiency and objectivity of data processing.

[0042] The working principle of this utility model is briefly described as follows: During the experiment, an appropriate amount of culture water was injected into the passageway 122, and zebrafish fry were introduced. According to the experimental design, movable partitions 123 were inserted or removed to configure the experimental space, and corresponding lighting programs were set. After the door 160 was closed, the device was activated. The lighting mechanism 140 provided the set lighting conditions, and the camera 150 began recording the entire process. The collected video data was transmitted in real time to the computer 170, where it was automatically analyzed by specialized software to ultimately obtain quantitative, objective, high-throughput experimental data on the social behavior of zebrafish fry.

[0043] The above description is merely a preferred embodiment of this application and an explanation of the technical principles and solutions employed. Furthermore, the scope of the utility model involved in this application is not limited to the specific combination of the above-described technical features, but should also cover other technical solutions formed by any combination of the above-described technical features or their equivalents without departing from the inventive concept. For example, technical solutions formed by substituting the above features with (but not limited to) technical features with similar functions disclosed in this application.

Claims

1. A zebrafish juvenile social behavior cognitive analysis device, including an analyzer body (100), characterized in that: The analyzer body (100) includes a housing (110), a passageway mechanism (120), a tray (130), a lighting mechanism (140), and a camera (150). The passageway mechanism (120) includes a passageway plate (121), which is detachably mounted on the top of the tray (130), and multiple independent passageway channels (122) are arranged side by side on the passageway plate (121). The lighting mechanism (140) is located below the tray (130) and is used to provide controllable lighting to the passageway (122); The camera (150) is located at the top inside the box (110) and directly above the passageway plate (121) for filming the behavior of zebrafish fry in the passageway channel (122).

2. The zebrafish juvenile social behavior cognitive analysis device according to claim 1, characterized in that: The number of the passageways (122) is ten, and each passageway (122) is a rectangular groove structure.

3. The zebrafish juvenile social behavior cognitive analysis device according to claim 2, characterized in that: At least one movable partition (123) is provided inside the passageway (122), which divides the passageway (122) into multiple experimental sections.

4. The zebrafish juvenile social behavior cognitive analysis device according to claim 3, characterized in that: The movable partition (123) is installed in the slot of the passageway plate (121) by snap-fit ​​or plug-in.

5. The zebrafish juvenile social behavior cognitive analysis device according to claim 1, characterized in that: A side panel (124) is provided between two adjacent passageways (122), and the side panel (124) is made of opaque material to achieve visual isolation.

6. The zebrafish juvenile social behavior cognitive analysis device according to claim 1, characterized in that: The lighting mechanism (140) includes a lighting panel (141) with multiple independent lighting areas (142) on the lighting panel (141), each of the lighting areas (142) being able to emit at least two different colors of light.

7. The zebrafish juvenile social behavior cognitive analysis device according to claim 6, characterized in that: The number of the lighting areas (142) is three, arranged side by side along the transverse direction of the lighting panel (141); each lighting area (142) can independently emit white, green, blue or red light.

8. The zebrafish juvenile social behavior cognitive analysis device according to claim 1, characterized in that: The front end of the box (110) is provided with an opening, and a door (160) that can be raised and lowered in the vertical direction is installed at the opening.

9. The zebrafish juvenile social behavior cognitive analysis device according to claim 1, characterized in that: The camera (150) is connected to an external computer (170) via a data cable. The computer (170) is equipped with analysis software for analyzing images of zebrafish behavior.

10. The zebrafish juvenile social behavior cognitive analysis device according to claim 1, characterized in that: The bottom of the tray (130) has a conical structure.

Images