Zebra fish behavior shooting system and method

Abstract

The invention discloses a zebra fish behavior shooting system and method, and relates to the technical field of image processing, and the system comprises a camera which is arranged at the top of a box body; the experiment module is used for culturing zebra fish, and a camera shoots images of the experiment module and the zebra fish; and the computer is used for detecting the edge of the experiment module in the image, taking an area enclosed by the edge as an activity area of the zebra fish, identifying the position of the zebra fish in the activity area, and connecting the positions to form a track diagram of the zebra fish. The application has remarkable advantages in compatibility, and can adapt to various different types of pore plates and self-defined experiment modules. Through flexible hardware configuration and modular design, the system can be customized and adjusted according to experiment requirements, limitation of traditional equipment during experiment switching is avoided, and larger experiment freedom degree and applicability are provided for researchers.

Description

Technical Field

[0001] This application relates to the field of image processing technology, and particularly relates to a zebrafish behavior shooting system and method. Background Art

[0002] As an important model organism, zebrafish has been widely used in the fields of neuroscience, ethology, toxicology, and drug screening due to its unique advantages of transparent embryos, rapid development, and easy genetic manipulation. Especially in ethological research, by analyzing the movement trajectories and behavior patterns of zebrafish, the pathological mechanisms of neurological diseases, the effects of drugs, and the impact of environmental factors on organisms can be deeply revealed. Therefore, developing a set of efficient and precise zebrafish behavior shooting and analysis system is of great significance for promoting the progress of behavioral biology and related research fields.

[0003] Currently, zebrafish behavior research equipment mainly uses traditional single-function devices, such as the method of combining high-resolution camera equipment with manual data analysis software. Some systems have basic behavior tracking functions and can record the movement trajectories of zebrafish and generate preliminary behavioral analysis data. Some devices also support simple environmental control functions, such as temperature and light adjustment, to reduce the interference of the external environment on experimental results. However, most of these systems have the deficiencies of single function and poor scalability, and it is difficult to meet the requirements of different experimental scenarios. For example, their adaptability to well plates or special-shaped experimental modules such as T-mazes and custom devices is limited. Summary of the Invention

[0004] The embodiments of this application provide a zebrafish behavior shooting system to solve the problem of poor adaptability to different experimental scenarios in the prior art.

[0005] On the one hand, the embodiments of this application provide a zebrafish behavior shooting system, including:

[0006] A box body;

[0007] A top cover, arranged on the top of the box body, and a camera facing the inside of the box body is arranged on the top cover;

[0008] A hot air box, arranged on the back of the box body, and the hot air box is used to generate hot air at a set temperature and convey the hot air to the inside of the box body;

[0009] A light source, arranged on the inner bottom surface of the box body, and the light source is used to generate a vertical upward surface light source;

[0010] An experimental module, used for culturing zebrafish, the experimental module is placed on the light source, and the camera shoots images of the experimental module and zebrafish;

[0011] A computer, electrically connected to a camera, is used to detect the edges of the experimental module in the image, take the area enclosed by the edges as the activity area of the zebrafish, identify the positions of the zebrafish in the activity area, and connect the positions in chronological order to form a trajectory map of the zebrafish.

[0012] On the other hand, an embodiment of the present application also provides a method for photographing zebrafish behavior, including:

[0013] Place the experimental module with zebrafish on the light source and turn on the light source;

[0014] Set the target temperature and start the hot air box. After heating the air to the target temperature, the hot air box transports it into the box where the experimental module is stored;

[0015] Connect the camera and the computer. The camera is set on the top of the box. The camera takes images of the experimental module and the zebrafish and transmits the images to the computer;

[0016] The computer detects the edges of the experimental module in the image, takes the area enclosed by the edges as the activity area of the zebrafish, identifies the positions of the zebrafish in the activity area, and connects the positions in chronological order to form a trajectory map of the zebrafish.

[0017] A zebrafish behavior photographing system and method in the present application have the following advantages:

[0018] 1. Existing zebrafish behavior research systems have the disadvantage of poor adaptability and cannot meet diverse experimental needs. The zebrafish behavior photographing system of the present application has significant advantages in compatibility and can adapt to various different types of well plates such as 1-well plate, 6-well plate, 24-well plate, and 96-well plate, as well as custom experimental modules such as T-maze and special structure designs. Through flexible hardware configuration and modular design, the system can be customized according to experimental needs, avoiding the limitations of traditional equipment during experimental switching, and providing greater experimental freedom and applicability for researchers.

[0019] 2. The zebrafish behavior photographing system of the present application performs excellently in environmental control. The built-in temperature control chip can precisely adjust the temperature of the experimental environment, and through sound insulation optimization design, it minimizes external interference. The main box adopts an efficient heat insulation and flow disturbance structure to ensure uniform temperature distribution inside the box, avoiding experimental errors caused by temperature fluctuations in traditional equipment. This environmental optimization ability not only ensures the normal physiological state of zebrafish but also improves the repeatability and reliability of experimental data.

[0020] 3. The zebrafish behavior recording system of the present application supports real-time video preview and visual display of behavior analysis results. Through a high-resolution camera and intelligent analysis algorithms, it can capture the behavior trajectories of zebrafish in real time and generate dynamic result graphs. The system can directly display the movement paths, area preferences, and key behavior characteristics of zebrafish during the experiment, significantly enhancing the visibility and interpretation efficiency of experimental data. Researchers can not only immediately adjust the experimental conditions but also quickly verify the experimental effects, providing a more intuitive and efficient research tool for zebrafish behavior research.

[0021] 4. The zebrafish behavior recording system of the present application integrates a high-resolution camera and intelligent analysis software, achieving the integrated function of shooting and analysis. Researchers can complete real-time capture, automated analysis, and data processing of zebrafish behavior on the same platform without relying on additional software or hardware support. Precise movement trajectory tracking and data classification analysis significantly improve the experimental efficiency and the reliability of results, while reducing human intervention and errors, providing an efficient, convenient, and intuitive full-process solution for zebrafish behavior research. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments of the present application. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0023] Figure 1 It is a schematic diagram of the closed state of the box body in the zebrafish behavior recording system provided by the embodiment of the present application.

[0024] Figure 2 It is a schematic diagram of the back structure of the box body provided by the embodiment of the present application.

[0025] Figure 3 It is a schematic diagram of the structure of the light source and the experimental module inside the box body provided by the embodiment of the present application.

[0026] Figure 4 It is a schematic diagram of the top cover and the camera thereon provided by the embodiment of the present application.

[0027] Figure 5 It is a schematic diagram of the open state of the box body provided by the embodiment of the present application.

[0028] Figure 6 They are images of various different experimental modules obtained by the camera provided by the embodiment of the present application.

[0029] Figure 7The trajectory diagram of zebrafish obtained by computer analysis provided by the embodiments of the present application.

[0030] Figure 8 The edge diagrams of various different well plates obtained by computer analysis provided by the embodiments of the present application.

[0031] Figure 9 The trajectory diagram of zebrafish in a 24-well plate obtained by computer analysis provided by the embodiments of the present application.

[0032] Explanation of the reference numerals in the drawings: 1. Main body box; 2. Box door; 3. Top cover; 4. Hot air box; 5. Fan; 6. Hot air pipe; 7. Temperature control unit; 8. Light source; 9. Light homogenizing plate; 10. Experimental module; 11. Camera; 12. Support plate. Detailed implementation manners

[0033] Next, the technical solutions in the embodiments of the present application will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present application.

[0034] Figures 1-5 A partial structural schematic diagram of a zebrafish behavior photographing system provided by the embodiments of the present application. The embodiments of the present application provide a zebrafish behavior photographing system, including:

[0035] Box body;

[0036] Top cover 3, arranged on the top of the box body, and a camera 11 facing the inside of the box body is arranged on the top cover 3;

[0037] Hot air box 4, arranged on the back of the box body, and the hot air box 4 is used to generate hot air at a set temperature and convey the hot air to the inside of the box body;

[0038] Light source 8, arranged on the inner bottom surface of the box body, and the light source 8 is used to generate a vertical upward surface light source;

[0039] Experimental module 10, used for culturing zebrafish, the experimental module 10 is placed on the light source 8, and the camera 11 photographs the experimental module 10 and the images of zebrafish;

[0040] Computer, electrically connected to the camera 11, and the computer is used to detect the edge of the experimental module 10 in the image, take the area enclosed by the edge as the activity area of the zebrafish, identify the position of the zebrafish in the activity area, and connect the positions in chronological order to form the trajectory diagram of the zebrafish.

[0041] Exemplarily, the box body includes a main box 1 and a box door 2. Both the main box 1 and the box door 2 are made of aluminum alloy. The main box 1 is a hollow cube structure with an opening on the front side. The box door 2 is rotatably connected to the position of the opening on the front side of the main box 1 through a hinge. When the box door 2 is closed, the opening can be completely sealed, reducing the adverse effects of the external environment on the internal environment of the main box 1.

[0042] On the top surface of the main box 1, there is an installation hole slightly smaller than the top cover 3. At the position of the main box 1's top surface near this installation hole, there are multiple screw holes, and on the top cover 3 near the edge, there are through holes matching the positions of the screw holes on the main box 1. When the top cover 3 covers the installation hole, the screw holes will correspond to the through holes one by one. At this time, the top cover 3 can be installed on the top surface of the main box 1 using bolts.

[0043] In the embodiment of the present application, the camera 11 is plugged into the support plate 12. There is also an installation hole on the top cover 3. At the position of the top cover 3 near this installation hole, there are multiple screw holes, and on the support plate 12 near the edge, there are through holes matching the positions of the screw holes on the top cover 3. When the support plate 12 covers the installation hole, the screw holes will correspond to the through holes one by one. At this time, the support plate 12 can be installed on the top cover 3 using bolts. On the side surface of the support plate 12 located inside the main box 1, there is a groove matching the shape and size of the camera 11, and the camera 11 is plugged into this groove. The cable connected to the camera 11 passes through the bottom of the groove and extends out of the main box 1.

[0044] Further, a light homogenizing plate 9 is arranged above the light source 8, and the experimental module 10 is placed on the light homogenizing plate 9. The size and shape of the light source 8 are equivalent to the cross-section of the internal space of the main box 1. Therefore, when the box door 2 is opened, the light source 8 installed at the bottom inside the main box 1 can completely occupy the bottom surface, so that the emitted surface light source can be distributed as evenly as possible inside the main box 1. The light homogenizing plate 9 can scatter the light emitted by the light source 8 in a specific direction in different directions to further improve the uniformity of the internal illumination of the main box 1.

[0045] Specifically, in the embodiment of the present application, a bracket is placed on the top surface of the light source 8. This bracket is formed by splicing multiple vertical thin plates, and the light homogenizing plate 9 is placed on the bracket. After setting the bracket, the distance between the light source 8 and the light homogenizing plate 9 can be increased, so that the divergent light emitted by the light source 8 can irradiate the entire light homogenizing plate 9, further enhancing the uniformity of the illumination.

[0046] It should be understood that regardless of the type, the experimental module 10 is made of a transparent material so that the light below can pass through the experimental module 10 and finally be received by the camera 11 to form an image of the experimental module 10 and the zebrafish.

[0047] Furthermore, a temperature control unit 7 is provided on the outer side surface of the main body box 1. The temperature control unit 7 is internally provided with a temperature control chip, and at the same time, an operation panel is provided on the outer surface of the temperature control unit 7. Researchers can set the target temperature through the operation panel, and after the temperature control chip obtains the target temperature, it can control the hot air box 4.

[0048] Specifically, the hot air box 4 is provided with an air inlet on the side surface facing away from the main body box 1. A fan 5 is provided in the air inlet. The fan 5 works under the control of the temperature control chip to transport the air outside the hot air box 4 into the hot air box 4. In addition, a heating unit is also provided inside the hot air box 4. The heating unit can be integrated with the fan 5. The heating unit generates heat under the control of the temperature control chip, so that the air transported by the fan 5 is heated to the target temperature.

[0049] In the embodiment of the present application, hot air pipes 6 are respectively connected to two side surfaces adjacent to the side surface of the hot air box 4 with the fan 5. The ends of the hot air pipes 6 are both connected to the main body box 1 to communicate the inside of the hot air box 4 and the main body box 1. Specifically, the inner diameter of the hot air pipe 6 is 100 mm, the width of the side surface of the hot air box 4 connected to the hot air pipe 6 is 145 mm, the distance between the fan 5 and the inner side surface of the hot air box 4 close to the main body box 1 is 125 mm, and both ends of the hot air pipe 6 are connected to the hot air box 4 and the main body box 1 through bolts.

[0050] The camera 11 in the embodiment of the present application uses a high-definition industrial camera module with 48 million pixels, supports multiple resolutions and PDAF phase focusing modes, and the operating temperature range is -20°C to 60°C. The working voltage of the light source 8 is 12 - 24V, the size of the light-emitting part is 442 * 416 mm, it has the characteristics of long life and no stroboscopic, and at the same time, it also has good heat dissipation ability.

[0051] In order to reduce the heat loss inside the main body box 1, the embodiment of the present application also mounts a heat insulation layer on the inner surface of the main body box 1. Moreover, since the hot air pipe 6 is connected to a fixed position of the main body box 1, the hot air will also be input into the main body box 1 from this fixed position. In order to avoid heat accumulation at the position close to the hot air pipe 6, the embodiment of the present application also provides a flow disturbance structure inside the main body box 1. The flow disturbance structure is a plurality of flow disturbance plates. These flow disturbance plates are generally fixed at the position inside the main body box 1 close to the hot air pipe 6, and the other ends are fixed at the position close to the experimental module 10. These flow disturbance plates are arranged in a parallel or at a certain angle to each other, so that the hot air input by the hot air pipe 6 is cut into multiple strands by the space between the plurality of flow disturbance plates, and the flow rate of each strand of hot air is basically the same. These hot air will be guided to various positions inside the main body box 1, so that the heat can be evenly dispersed in the main body box 1, providing a good environment for the activities of zebrafish.

[0052] In a possible embodiment, the experimental module 10 is one of a well plate and a special-shaped module. The computer obtains the module type selected by the researcher and detects the edge according to the module type. When the experimental module 10 is a well plate, the computer detects the inner edge of the well plate and takes the area enclosed by the inner edge as the active area. When the experimental module 10 is a special-shaped module, the computer detects the outer edge of the special-shaped module and takes the area enclosed by the outer edge as the active area.

[0053] Exemplarily, the well plate is a single-well plate or a multi-well plate. The single-well plate is a culture dish, and the multi-well plate is provided with a plurality of circular or oval culture grooves in a multi-row and multi-column manner on a rectangular bottom plate. The zebrafish are cultured in the culture dish or culture grooves.

[0054] Before the experiment, the researcher needs to measure the parameters of the experimental module 10 in advance, and then define the characteristic information of the experimental module 10 in the intelligent analysis software of the computer. The characteristic information includes the module type and the parameters under each module type. After the experimental module 10 is placed in the main body box 1, the researcher needs to select the module type of the experimental module 10 in the intelligent analysis software. The module type includes a well plate and a special-shaped module. If a well plate is selected, the number of holes on the well plate also needs to be selected, such as 1 hole, 6 holes, 24 holes, 96 holes, etc. If a special-shaped module is selected, the specific shape also needs to be selected, including a T shape and other shapes. After selecting the specific module type, the computer will detect the edge of the experimental module 10 according to the parameters corresponding to the module type.

[0055] Further, before the camera 11 starts to collect images, the researcher needs to set the time interval and the image saving path in the intelligent analysis software. After starting to shoot, the camera 11 will shoot images at the set time interval and save the taken images in a specific location. The images taken by the camera 11 in the embodiment of the present application are as Figure 6 shown, and the trajectory diagram analyzed by the computer is as Figure 7 shown.

[0056] Further, the method for the computer to detect the inner edge includes: S1, converting the color image into a corresponding grayscale image; S2, segmenting the grayscale image into foreground and background according to the segmentation threshold, and converting the foreground and background into black and white respectively to obtain a binary image; S3, analyzing the gradient of the binary image and determining the edge points according to the gradient; S4, using the convex hull algorithm to connect the edge points in sequence to form a plurality of convex hulls; S5, performing shape analysis on the convex hulls to determine the inscribed circle or inscribed ellipse in each convex hull; S6, filtering the plurality of inscribed circles or inscribed ellipses, and the finally obtained inscribed circle or inscribed ellipse is the inner edge.

[0057] Specifically, the image received by the computer is as Figure 6As shown, these images may contain some color information. To reduce the computational load of the computer, the present application first converts the images into grayscale images according to the following formula:

[0058] Gray = 0.2989R + 0.5870G + 0.1140B

[0059] Where Gray represents the grayscale value of each pixel in the grayscale image, and R, G, and B represent the values of each pixel in the red, green, and blue channels respectively.

[0060] Using the above formula to compress the data of the three channels into one channel reduces the amount of data and the computational complexity. After removing the color information while retaining the key information, the key information such as the shape, edges, and texture of the object can still be retained, which provides a basis for subsequent edge detection and edge extraction and other processes.

[0061] In step S2, the computer establishes a neighborhood window centered on each pixel in the grayscale image, determines the segmentation threshold of the central pixel according to the mean value of all pixels in the neighborhood window, and performs foreground and background segmentation on each pixel according to the segmentation threshold.

[0062] Specifically, through adaptive segmentation threshold calculation, the threshold of each pixel in the grayscale image is dynamically determined, and the segmentation threshold is generated by considering the grayscale distribution of the local neighborhood. The formula is:

[0063] T(i,j) = Mean(R(i,j)) - C

[0064] Where T(i,j) represents the segmentation threshold of the pixel at position (i,j), R(i,j) represents the neighborhood window centered on the pixel (i,j), Mean(R(i,j)) represents the average value of the grayscale values of all pixels in the neighborhood window, and C is a constant used to adjust the local threshold to reduce background misdetection. By dynamically generating the segmentation threshold for each pixel in the embodiments of the present application, the problems of illumination change and complex background can be processed more effectively, and the segmentation errors caused by the global fixed segmentation threshold can be avoided.

[0065] In the binarization operation of step S2, the grayscale value of the pixel in the grayscale image is compared with the segmentation threshold, and the pixel is classified as foreground or background according to the comparison result, and the grayscale image is converted into a binary image containing only black and white. The formula is:

[0066]

[0067] Where B(i,j) represents the value after the binarization process of the pixel (i,j), the value of 1 indicates classification as background, and the pixel will be converted to white, and the value of 0 indicates classification as foreground, and the pixel will be converted to black, and I(i,j) represents the grayscale value of the pixel (i,j).

[0068] After converting the grayscale image into a binary image, the data structure is simplified, facilitating the detection and analysis of the target, thereby clearly distinguishing the target, that is, the foreground and the background.

[0069] After step S2, for the obtained binary image, morphological denoising processing is also performed to filter the connected regions in the binary image, and then gradient analysis is performed on the processed binary image.

[0070] Specifically, by removing small connected regions in the binary image caused by noise or non-target regions, the purity of the target region is enhanced. This application adopts connected region filtering based on area. The formula is:

[0071] Area(Rk)<A min →Rk is removed

[0072] where Rk represents the k-th connected region in the binary image, Area(Rk) represents the pixel area of the connected region Rk, and A min represents the minimum area threshold set by the user. Connected regions below this value will be removed. By removing the noise region, this application effectively reduces false detections, highlights the real target region, significantly improves the quality of the binary image, and provides more reliable data support for subsequent morphological analysis and feature extraction.

[0073] In step S3, the core of edge detection is to identify positions in the image where the pixel intensity changes significantly, which usually correspond to the boundaries of objects. By calculating the gradient of the grayscale value in the image (i.e., the grayscale change rate), edge points can be detected. These edge points are connected together to form the boundary of the object. The gradient is the rate of change of the grayscale value with position, and significant grayscale changes usually occur at the edges. Therefore, the gradient magnitude is an important basis for judging edges. In the process of calculating the gradient in step S3, first calculate the magnitude of the gradient. The magnitude G of the image gradient is calculated by the Sobel operator:

[0074]

[0075] where G represents the gradient magnitude of the pixel point, which reflects the intensity change of the pixel point, and respectively represent the change rates of the grayscale value I of the image in the x and y directions.

[0076] Then perform direction calculation. The direction θ of the gradient represents the direction of the edge:

[0077]

[0078] Finally, according to the set gradient threshold, the pixels greater than the threshold are retained as edge points. By extracting the boundary information of the object, it provides a basis for contour extraction. In complex backgrounds, it can effectively separate the target area from the background area.

[0079] In step S4, the convex hull is the smallest convex polygon of the target area, which is a convex set containing all edge points of the target. The advantage of the convex hull algorithm is that it can quickly generate the overall shape of the target area, which is suitable for describing the boundaries of regular and most irregular targets. The principle is that given a set of points S, the convex hull H is a smallest convex polygon containing all points, satisfying:

[0080]

[0081] Among them, H represents the minimum polygon containing the edge point set S of the target area, P i , P j and P k Represent the i-th, j-th and k-th edge points respectively.

[0082] After all edge points are obtained in step S3, the edge points are scanned step by step in the order of polar angle from small to large and connected in sequence to form a convex hull. The convex hull provides a geometric description of the target area, which is particularly suitable for the boundary representation of regular or approximately regular shapes, and can quickly determine the shape of the target area for morphological analysis or target detection.

[0083] For the experimental module 10 in the form of a well plate, the holes thereon may be circular or elliptical, and the embodiment of the present application takes circular holes as an example.

[0084] The core of circular hole detection is to use image processing technology and geometric analysis methods to extract the area that meets the circular characteristics. In the analysis process, the convex hull is analyzed by circular Hough transform to determine whether the convex hull meets the circular characteristics. The convex hull that meets the circular characteristics is retained, and the inscribed circle is determined in the retained convex hull. The standard equation of a circle is:

[0085] xx c 2 +yy c 2 =r 2

[0086] Among them, (x c ,y c ) is the coordinate of the center of the circle, and r is the radius of the circle. Each pixel in the image is verified by this equation to see if it meets the circular feature.

[0087] Then use the circular Hough transform to detect the circle and find the peak of the cumulative number of points of the circle in the parameter space:

[0088]

[0089] Among them, H(x c , y c , r) represents the number of votes in the parameter space, (x, y) represents the coordinates of the edge point, Edge represents the set of edge points, and δ is the unit impulse function used to verify whether the pixel point conforms to the circular equation.

[0090] For multiple convex hulls established, the convex hulls that conform to the circular characteristic equation will be retained, while those that do not conform will be deleted. Among the retained convex hulls, the inscribed circles will continue to be drawn to determine the inner edge of the hole in the experimental module 10.

[0091] After drawing the inscribed circles, the inscribed circles can be filtered according to the pre-set minimum radius threshold, maximum radius threshold, sensitivity parameter, and distance threshold. Among them, when screening according to the distance threshold, by calculating the relationship between the distance between the centers of the circles and the radius, the circles that are too close are removed:

[0092] d(C i , C j ) < R min → Remove the smaller circle

[0093] Among them, d(C i , C j ) is the distance between the center of the circle C i and the center of the circle C j , and R min is the minimum radius threshold.

[0094] Through the overlapping filtering algorithm, redundant circles are removed to ensure the detection accuracy. The detection result of the inner edge can be visually displayed on the original image, as shown in Figure 8 , providing intuitive feedback to the user for easy verification and adjustment of parameters. The trajectory analysis result of zebrafish in each hole when using a 24-well plate is as shown in Figure 9 .

[0095] This embodiment of the present application also provides a method for photographing zebrafish behavior, and this method includes the following steps:

[0096] Place the experimental module 10 with cultured zebrafish on the light source 8 and turn on the light source 8;

[0097] Set the target temperature and start the hot air box 4. After heating the air to the target temperature, the hot air box 4 conveys it to the box where the experimental module 10 is placed;

[0098] Connect the camera 11 and the computer. The camera 11 is set on the top of the box, and the camera 11 takes pictures of the experimental module 10 and the zebrafish and transmits the pictures to the computer;

[0099] The computer detects the edges of the experimental module 10 in the image, takes the area enclosed by the edges as the activity area of the zebrafish, identifies the positions of the zebrafish in the activity area, and connects the positions in chronological order to form a trajectory map of the zebrafish.

[0100] Although the preferred embodiments of the present application have been described, those skilled in the art can make additional changes and modifications to these embodiments once they learn the basic creative concepts. Therefore, the appended claims are intended to be construed as including the preferred embodiments as well as all changes and modifications that fall within the scope of the present application.

[0101] Obviously, those skilled in the art can make various changes and modifications to the present application without departing from the spirit and scope of the present application. Thus, if these modifications and variations of the present application fall within the scope of the claims of the present application and their equivalent technologies, the present application is also intended to include these modifications and variations.

Claims

1. A zebrafish behavior filming system, characterized in that: include: Box; A top cover (3) is arranged on the top of the box body, and a camera (11) facing the inside of the box body is arranged on the top cover (3); A hot air box (4) is arranged at the back of the box, and the hot air box (4) is used to generate hot air of a set temperature and transport the hot air to the inside of the box; A light source (8) is arranged on the inner bottom surface of the box, and the light source (8) is used to generate a surface light source pointing vertically upward; An experimental module (10) for cultivating zebrafish, wherein the experimental module (10) is placed on the light source (8), and the camera (11) captures images of the experimental module (10) and the zebrafish; A computer is electrically connected to the camera (11), and the computer is used to detect the edge of the experimental module (10) in the image, use the area enclosed by the edge as the activity area of ​​the zebrafish, identify the position of the zebrafish in the activity area, and connect the positions in chronological order to form a trajectory map of the zebrafish.

2. A zebrafish behavior filming system according to claim 1, characterized in that: The experimental module (10) is one of a hole plate and a special-shaped module. The computer obtains the module type selected by the researcher and detects the edge according to the module type. When the experimental module (10) is a hole plate, the computer detects the inner edge of the hole plate and uses the area enclosed by the inner edge as the active area. When the experimental module (10) is a special-shaped module, the computer detects the outer edge of the special-shaped module and uses the area enclosed by the outer edge as the active area.

3. A zebrafish behavior filming system according to claim 2, characterized in that: The method for detecting the inner edge by a computer comprises: Converting the color image into a corresponding grayscale image; Segmenting the grayscale image into a foreground and a background according to a segmentation threshold, converting the foreground and the background into black and white respectively, to obtain a binary image; Analyzing the gradient of the binary image, and determining edge points according to the gradient; Using a convex hull algorithm to connect the edge points in sequence to form a plurality of convex hulls; Performing shape analysis on the convex hull to determine an inscribed circle or an inscribed ellipse in each of the convex hulls; The multiple inscribed circles or inscribed ellipses are filtered, and the inscribed circle or inscribed ellipse finally obtained is the inner edge.

4. A zebrafish behavior filming system according to claim 3, characterized in that: The computer establishes a neighborhood window with each pixel in the grayscale image as the center, determines the segmentation threshold of the central pixel according to the mean value of all pixels in the neighborhood window, and performs foreground and background segmentation on each pixel according to the segmentation threshold.

5. A zebrafish behavior filming system according to claim 3, characterized in that: After the binary image is obtained, a morphological denoising process is performed on the binary image to filter the connected areas in the binary image, and the processed binary image is then subjected to a gradient analysis.

6. A zebrafish behavior filming system according to claim 3, characterized in that: The plurality of edge points are connected in sequence in order of polar angle from small to large to form the convex hull.

7. A zebrafish behavior filming system according to claim 3, characterized in that: The convex hull is subjected to shape analysis by using Hough transform to determine whether the convex hull meets the characteristics of a circle or an ellipse, the convex hull that meets the characteristics of a circle or an ellipse is retained, and the inscribed circle or the inscribed ellipse is determined in the retained convex hull.

8. A zebrafish behavior filming system according to claim 7, characterized in that: The multiple inscribed circles or inscribed ellipses are filtered according to a preset radius threshold and a distance threshold.

9. A zebrafish behavior filming system according to claim 1, characterized in that: A light-diffusing plate (9) is arranged above the light source (8), and the experimental module (10) is placed on the light-diffusing plate (9).

10. A method for applying a zebrafish behavior filming system as claimed in any one of claims 1 to 9, characterized in that: include: Placing the experimental module (10) in which zebrafish are cultured on the light source (8), and turning on the light source (8); Setting a target temperature and starting a hot air box (4), wherein the hot air box (4) heats the air to the target temperature and then delivers the air to a box storing the experimental module (10); Connecting a camera (11) and a computer, wherein the camera (11) is arranged on the top of the box, and the camera (11) takes images of the experimental module (10) and the zebrafish, and transmits the images to the computer; The computer detects the edge of the experimental module (10) in the image, uses the area enclosed by the edge as the activity area of ​​the zebrafish, identifies the position of the zebrafish in the activity area, and connects the positions in chronological order to form a trajectory map of the zebrafish.

Images