Device for zebrafish juvenile fish light and shade preference behavior experiment

Abstract

The utility model discloses a device for a zebra fish juvenile fish light and shade preference behavior experiment, and relates to the technical field of zebra fish behavioristics research. Comprising a bottom plate and experiment units arranged on the bottom plate, the experiment units are 24 test holes which are formed in the bottom plate in four transverse rows and six longitudinal columns, the experiment units are arranged on the front faces of the 24 test holes through infrared light-transmitting black optical filters, dark areas are formed in the test holes through coated infrared light-transmitting black ink, and the infrared light-transmitting black optical filters are arranged on the front faces of the 24 test holes. And the dark area and the bright area are presented in the same test hole. After a video of the movement behavior of the juvenile fish is shot, behavioral analysis software can be used for analyzing the preferences of the bright area and the dark area of the juvenile fish, and whether the juvenile fish has neurotoxic effects such as anxiety behaviors or not is judged. According to the technical scheme, the dark area can be constructed without influencing shooting of the infrared camera, and the black ink transmitting infrared light avoids the painting mode of intervals between the holes, so that the ink is prevented from forming gathered dark color blocks, the movement behavior of the juvenile fish cannot be misjudged by behavioral analysis software due to the dark color blocks, and the operation is rapid and convenient.

Description

TECHNICAL FIELD

[0001] The utility model relates to zebra fish ethology research technical field especially, it relates to a device for zebra fish fry light and dark preference behavior experiment. BACKGROUND

[0002] The types of pollutants in the environment are increasing, which not only pollute water sources, but also can directly or indirectly affect humans through water, soil, air and other means. These pollutants can cause neurotoxic effects on the human body. In the human body, the nervous system is closely related to other organs and systems. When environmental pollutants have neurotoxic effects on the nervous system, the normal activities of the human body will be disturbed, and even diseases will occur.

[0003] Zebra fish is a small tropical fish, its genes have 87% high homology with human genes, has short breeding cycle and is easy to raise, and has low breeding cost. In addition, zebra fish is often used for water quality environmental monitoring and research on some environmental toxicant toxicity and mechanism of action as a model organism under laboratory conditions. Zebra fish is sensitive to pollutant toxicity in the embryonic development stage, and the experimental cycle is short, and the changes of nerve cells are easy to be characterized by existing technology, therefore, zebra fish fry is widely used in toxicology research.

[0004] Zebra fish fry is sensitive to external stimulation and adjusts behavior accordingly, and the motor behavior is regulated by the nervous system, therefore, the behavior changes of zebra fish fry are often used to evaluate the neurotoxic effects of environmental pollutants in the laboratory. In ethology research, the anxiety-like behavior of zebra fish fry is often evaluated by light and dark preference experiment. The behaviors that zebra fish fry stays in the dark environment for a longer time and the total swimming distance in the dark environment increases can indicate that the anxiety level of zebra fish fry increases. In the light and dark preference experiment of zebra fish fry, researchers usually use infrared cameras or behavior analysis instruments to record the motor behavior of zebra fish, and then use behavior analysis software to analyze the residence time, swimming distance and other indicators in the light and dark areas, further judge whether the zebra fish has neurotoxic effects and whether the effects are related to anxiety-like behavior, and evaluate the environmental pollution condition accordingly.

[0005] Most of the light and dark preference experiments are carried out in the light cycle of zebra fish fry, and a lot of data need to be recorded in the behavior experiment. The device for light and dark preference experiment of zebra fish fry is easy to have a dark color block due to incorrect design of the dark area, the dark color block will interfere with the recording of the movement track of the fry by the behavior analysis software, also cause calculation errors of the movement distance, movement speed and other indicators, and waste valuable experimental time. UTILITY MODEL CONTENT

[0006] The utility model discloses a device for zebra fish juvenile light-dark preference behavior experiment, which can solve the problem that dark color blocks in the dark area are easily misjudged by behavior analysis software in the zebra fish juvenile light-dark preference behavior experiment.

[0007] To achieve the above purpose, the technical scheme of the utility model is:

[0008] A device for zebra fish juvenile light-dark preference behavior experiment, comprising an experimental device body, which comprises a bottom plate and an experimental unit provided on the bottom plate, wherein the experimental unit is 24 test holes arranged in four horizontal rows and six vertical columns on the bottom plate, the 24 test holes have the same aperture, and adjacent test holes are connected by a ridge.

[0009] The utility model discloses a device for zebra fish juvenile light-dark preference behavior experiment, which can solve the problem that dark color blocks in the dark area are easily misjudged by behavior analysis software in the zebra fish juvenile light-dark preference behavior experiment.

[0010] Further, the ridge at the connection between the test holes and the test holes, and the connection between the test holes and the bottom plate are not coated with the infrared light transparent black ink, and the distance between the edge of the coated infrared light transparent black ink and the ridge is 2 mm.

[0011] Furthermore, the infrared-transmitting black filter is fixed to both sides of the base plate with transparent tape.

[0012] Furthermore, the test holes are evenly distributed on the base plate.

[0013] Furthermore, the test hole is circular or square.

[0014] The advantages and beneficial effects of this utility model are as follows:

[0015] 1. The device presents both light and dark areas in the same test hole, making it easy to capture the preference of juvenile fish for light and dark areas during their movement.

[0016] 2. The infrared-transmitting black filter is of moderate size and is fixed with transparent tape. This creates a dark area without affecting the infrared camera's recording. Furthermore, the infrared ink is applied in a way that avoids the gaps between the holes, preventing the ink from forming dark patches that could be misinterpreted by behavioral analysis software. This is the biggest highlight of this device.

[0017] 3. After the experiment, simply replace the original protective cap with the 24-well plate to store this invention. This protects the experimental materials and allows for convenient and environmentally friendly storage. The infrared-transmitting filter can be adjusted according to actual experimental needs, can be removed from the device after use, and can be reused, making it convenient and environmentally friendly. Attached Figure Description

[0018] Figure 1 This is a three-dimensional schematic diagram of the present invention.

[0019] Figure 2 This is a top view of the original 24-well plate.

[0020] Figure 3 This is a top view of the back of the original 24-hole plate.

[0021] Figure 4 This is a three-dimensional front view of the original 24-well plate (near the first column).

[0022] Figure 5 This is a three-dimensional front view of the original 24-well plate (near the 6th column).

[0023] Figure 6 This is a three-dimensional schematic diagram of the back of the original 24-well plate (near column 1).

[0024] Figure 7 This is a three-dimensional schematic diagram of the back of the original 24-well plate (near the 6th column).

[0025] Figure 8 This is a detailed perspective view of the front of the present invention (near column 1).

[0026] Figure 9 It is the stereogram of the front face of the utility model (close to column 6).

[0027] Figure 10 It is the stereogram of the front face of the utility model after removing the infrared light transparent black filter (close to column 1).

[0028] Figure 11 It is the stereogram of the front face of the utility model after removing the infrared light transparent black filter (close to column 6).

[0029] Figure 12 It is the stereogram of the back face of the utility model (close to column 1).

[0030] Figure 13 It is the stereogram of the back face of the utility model (close to column 6).

[0031] Figure 14 It is the schematic diagram of the infrared light transparent black filter of the utility model.

[0032] Figure 15 It is the front view of the utility model.

[0033] Figure 16 It is the front view of the utility model after removing the infrared light transparent black filter.

[0034] Figure 17 It is the back view of the utility model.

[0035] Figure 18 It is the shooting diagram of zebra fish larvae by Noldus animal motion trajectory tracking system and EthoVision XT 11.5 software in the embodiment.

[0036] Figure 19 It is the behavioral trajectory diagram of zebra fish larvae derived by EthoVision XT 11.5 software in the embodiment.

[0037] In the figure, 101. test hole, 102. infrared light transparent black filter, 103. transparent tape, 104. infrared light transparent black ink, 105. bottom plate. DETAILED DESCRIPTION

[0038] In order to make the technical problems, technical schemes and advantages of the utility model more clear, the following will be described in detail in combination with the drawings, specific implementation process and application scope.

[0039] The utility model provides a device for zebra fish fry light and dark preference behavior experiment, including bottom plate 105, and the experimental unit that is equipped with on bottom plate 105, wherein, the experimental unit is the 24 test holes 101 of setting on the bottom plate 105 horizontal four rows and vertical six columns, 24 test holes 101 have the same aperture, and equidistantly distribute on the bottom plate 105. The test hole of adjacent of 24 test holes is connected with test hole through the edge, still include infrared light black filter 102, this infrared light black filter 102 sets up on the front opening of 24 test holes 101 and forms dark area, infrared light black filter 102 is two, one infrared light black filter sets up in the area surrounded by the lower half of horizontal first row test hole center line and the upper half of second row test hole center line, and the other sets up in the area surrounded by the lower half of horizontal third row test hole center line and the upper half of fourth row test hole center line, the back of 24 test holes the lower half of horizontal first row test hole center line, with the upper half of second row test hole center line, and the lower half of horizontal third row test hole center line, with the upper half of fourth row test hole center line, surrounded area, are coated with infrared light black ink 104, the front of 24 test holes the lower half of horizontal first row test hole center line, with the upper half of second row test hole center line, surrounded area, the test hole outer wall of the test hole of the lower half of horizontal third row test hole center line, with the upper half of fourth row test hole center line, surrounded area, is coated with infrared light black ink 104.

[0040] The edge of the test hole and the test hole connection place and the test hole and the bottom plate connection place are not coated with infrared light black ink, and the distance between the edge of the coated infrared light black ink and the edge is 2mm. The infrared light black filter is fixed on both sides of the bottom plate by transparent adhesive tape. The test holes are equidistantly distributed on the bottom plate.

[0041] In the utility model, the drawings and the embodiments are only used for disclosing the device content, and are used for the person who needs and is familiar with the technology to understand, and the material model used in the device and the specific position of different components installation can be adjusted according to the need.

[0042] In the following embodiments, the experimental device body with 24 test holes 101 can use the 24-hole cell culture plate of the 702001 model of the NEST company, and the infrared light black ink 104 uses the water-insoluble black ink in the 6824-12 model marker pen of the Deli company. The infrared light black filter 102 can use the filter that meets the experimental requirements according to the experimental needs. If other brands and models of materials are replaced according to the actual experimental needs, as long as the purpose of the utility model can be achieved, and the expected technical effect of the utility model can be achieved.

[0043] The utility model experimental device is described first in combination with the drawings. Figure 1 The utility model discloses a device for zebra fish juvenile light-dark preference behavior experiment, which is composed of a body of the experimental device, infrared light transparent black filter 102, transparent adhesive tape 103 and infrared light transparent black ink 104. The body of the experimental device is composed of a bottom plate 105 and test holes 101. In this embodiment, the body of the experimental device is a 24-hole plate. Figure 2 As shown in the figure, the test hole at the upper left corner of the notch is A1 hole, and the hole is located in A row. From top to bottom, there are four rows of A, B, C and D, each row having six test holes. The column where A1 hole is located is column 1, and from left to right, there are six columns of 1, 2, 3, 4, 5 and 6, each column having four test holes. The test hole in the first row and the first column is marked as A1, the test hole in the first row and the second column is marked as A2, the test hole in the first row and the third column is marked as A3, the test hole in the first row and the fourth column is marked as A4, the test hole in the first row and the fifth column is marked as A5, and the test hole in the first row and the sixth column is marked as A6. In turn, the test hole in the second row and the first column is marked as B1, the test hole in the second row and the second column is marked as B2, the test hole in the second row and the third column is marked as B3, and so on, until the test hole in the fourth row and the sixth column is marked as D6, a total of 24 test holes. All the test holes 101 are evenly distributed on the bottom plate 105 as experimental units. Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 The figures are schematic diagrams of the original 24-hole plate.

[0044] Figure 8 Figure 9 The figures are schematic diagrams of the original 24-hole plate. Figure 15 Figure 8 Figure 9 The utility model infrared light transparent black filter 102 has two pieces, which are fixed above the test holes 101 by the transparent adhesive tape 103. One piece of filter covers the area surrounded by the lower half of the center line of the test holes in row A and the upper half of the center line of the test holes in row B. The other piece covers the area surrounded by the lower half of the center line of the test holes in row C and the upper half of the center line of the test holes in row D. Figure 4 Figure 5 The processing position is more characteristic in comparison. Figure 14 As shown in the figure, the distance from the center line of row A to the center line of row B is the width of the filter, and the length of the filter is equal to the length of the square frame on the front of the hole plate.

[0045] ​​​​​​​​The structure of this utility model after removing the infrared-transmitting black filter 102 is shown in the top view as follows. Figure 16 As shown, the 3D diagram is as follows Figure 10 , Figure 11 As shown. Figure 10 , Figure 11 The lower half of the center line of the test holes in row A, and the outer wall of the area enclosed by the upper half of the center line of the test holes in row B, are coated with infrared-transmitting black ink 104. The lower half of the center line of the test holes in row C, and the outer wall of the area enclosed by the upper half of the center line of the test holes in row D are coated with infrared-transmitting black ink 104. The slender connecting plate at the junction of the sidewall of the test hole and the sidewall of the adjacent test hole is not coated with the infrared-transmitting black ink 104, nor is the junction of the test hole and the base plate 105 coated with the infrared-transmitting black ink 104, and the edge of the infrared-transmitting black ink 104 is 2 mm away from the junction of the test hole and the junction of the test hole and the base plate 105. Figure 4 , Figure 5 Comparisons better reveal the characteristics of the processing location.

[0046] The top view of the back of the perforated plate of this utility model is as follows: Figure 17 As shown, the 3D diagram is as follows Figure 12 , Figure 13 As shown. Figure 17 On the bottom surface of the orifice plate, the lower half of the center line of row A test holes and the upper half of the center line of row B test holes form an area coated with infrared-transmitting black ink 104. The lower half of the center line of row C test holes and the upper half of the center line of row D test holes also form an area coated with infrared-transmitting black ink 104. Figure 3 Comparisons better reveal the characteristics of the processing location. Specifically, for example... Figure 12 , Figure 13 The outer walls of test holes A1, A6, D1, and D6 are coated with infrared-transmitting black ink 104. The elongated connecting plates at the junctions of test holes A1 and B1, C1 and D1, A6 and B6, and C6 and D6 are not coated with this infrared-transmitting black ink. The junctions of test holes A1, B1, C1, D1, A6, B6, C6, and D6 with the base plate 105 are also not coated with this infrared-transmitting black ink, and the edge of the infrared-transmitting black ink 104 is 2 mm away from the junctions between test holes and between the test holes and the base plate 105. Figure 12 , Figure 13 Enlarged views of test holes A1, D1, A6, and D6 are shown. Figure 6 , Figure 7 Comparisons better reveal the characteristics of the processing location.

[0047] The infrared light transparent black ink 104 of the outer side wall of the test hole and the outer bottom and the infrared light transparent black filter 102 above the test hole can construct a dark area environment, the infrared light transparent black ink 104 and the infrared light transparent black filter 102 used can transmit infrared light, and do not interfere with the shooting of the infrared camera on the movement behavior of the juvenile fish. The coloring mode of the infrared light transparent black ink 104 and the fixing mode of the infrared light transparent black filter 102 used in the utility model do not produce dark color blocks that affect the identification of the ethology analysis software, and are beneficial to the ethology analysis software to better analyze the light and dark preference of the juvenile fish.

[0048] The utility model takes using Noldus animal movement trajectory tracking system and EthoVision XT 11.5 software to carry out zebra fish juvenile light and dark preference experiment and movement trajectory identification as an example. After ensuring that the infrared light transparent black filter 102 is fixed firmly by the transparent adhesive tape 103 of the both sides of the hole plate, the Pasteur pipette is used to transfer the zebra fish juvenile into the test hole 101 of the utility model, after the transfer is finished, the utility model is put into the ethology recorder (the original 24 hole plate cover does not need to be covered), and the movement behavior of the zebra fish juvenile can be shot. After the test is finished, the hole plate can be taken out. After the juvenile is removed from the test hole, the utility model can be cleaned, and then the original 24 hole plate cover can be covered to be stored. Figure 18 The shooting effect diagram of the zebra fish juvenile by the Noldus animal movement trajectory tracking system and EthoVision XT 11.5 software is as shown in Figure 19 It can be seen that the color block that interferes with the ethology analysis software does not appear in the test hole 101, and the trajectory diagram does not appear the characteristics of the ethology software misjudgment such as long straight line, which shows that the utility model has strong practicability.

[0049] Based on the above embodiment, the utility model is mainly used for the light and dark preference behavior experiment of the zebra fish juvenile or other fish juvenile. After the juvenile is put into the test hole of the utility model, the light and dark preference behavior of the juvenile can be recorded according to the experiment arrangement, and then the preference of the juvenile to the light area and the dark area is analyzed, and whether the juvenile appears the anxiety-like behavior and other neurotoxicity effects is judged. The present embodiment is explained by using the 24 hole plate, and the purpose of the utility model can be achieved by using only the 24 hole plate. The test hole can be any aperture and depth of the top opening containing space, and the test hole can be circular or square or any polygon, and the utility model is not limited.

[0050] The utility model has simple structure but rich functions, and can be applied to the following several situations.

[0051] 1. When the infrared camera or the ethology recording instrument is used to shoot the light and dark preference of the juvenile in the laboratory, the utility model can be used as a device for constructing the light area and the dark area.

[0052] 2. Determine whether the experimental treatment of the juvenile fish has any neurotoxic effects such as anxiety-like behavior.

[0053] 3. After the infrared light transparent black filter 102 is removed, it can be used for preliminary judgment of whether the juvenile fish in the experiment has anxiety-like behavior such as light-dark preference. In the formal experiment, only the infrared light transparent black filter 102 is fixed on the original position of the filter with transparent tape 103, and the experiment can start, saving experimental materials and being convenient and fast.

[0054] The technical concept and technical scheme of the present application are described in detail through the above embodiments. It should be further pointed out that the present application has a simple structure and outstanding technical effects. The above embodiments are only used to understand the technical scheme of the present application, and the directions such as up, down, left and right involved in the embodiments are used to better understand the technical scheme of the present application, and are not used to limit the protection scope of the present application. The structures, shapes, materials and connection modes not specifically described in the present application all belong to the common knowledge in the art, and can be replaced by equivalent technologies in the related art. At the same time, any obvious adjustment and change to the structure and shape of the technical concept of the present application should also belong to the protection scope of the present application.

Claims

1. An apparatus for experimenting on light-dark preference behavior in juvenile zebrafish, characterized in that: The experimental device includes a base plate and an experimental unit on the base plate. The experimental unit consists of 24 test holes arranged in four horizontal rows and six vertical columns on the base plate. The 24 test holes have the same aperture, and adjacent test holes are connected by ridges. The system includes infrared-transmitting black filters, which are disposed on the front of 24 test holes. Two infrared-transmitting black filters are used: one is disposed in the area enclosed by the lower half of the center line of the first horizontal row of test holes and the upper half of the center line of the second horizontal row of test holes; the other is disposed in the area enclosed by the lower half of the center line of the third horizontal row of test holes and the upper half of the center line of the fourth horizontal row of test holes. The areas enclosed by the lower half of the center line of the first horizontal row of test holes and the upper half of the center line of the second horizontal row of test holes, as well as the areas enclosed by the lower half of the center line of the third horizontal row of test holes and the upper half of the center line of the fourth horizontal row of test holes, on the back of the 24 test holes are coated with infrared-transmitting black ink. The outer walls of the test holes in the areas enclosed by the lower half of the center line of the first horizontal row of test holes and the upper half of the center line of the second horizontal row of test holes on the front of the 24 test holes are coated with infrared-transmitting black ink, as are the outer walls of the test holes in the areas enclosed by the lower half of the center line of the third horizontal row of test holes and the upper half of the center line of the fourth horizontal row of test holes.

2. The apparatus for experimenting on light and dark preference behavior in juvenile zebrafish according to claim 1, characterized in that: The edges at the connection between the test holes and the connection between the test holes and the base plate are not coated with infrared-transmitting black ink, and the distance between the edge of the coated infrared-transmitting black ink and the edge is 2mm.

3. The apparatus for experimenting on light and dark preference behavior in juvenile zebrafish according to claim 1, characterized in that: The infrared-transmitting black filter is fixed to both sides of the base plate with transparent tape.

4. The apparatus for experimenting on light and dark preference behavior in juvenile zebrafish according to claim 1, characterized in that: The test holes are evenly distributed on the base plate.

5. The apparatus for experimenting on light and dark preference behavior in juvenile zebrafish according to claim 1, characterized in that: The test hole is circular or square.

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