| Summary: | 碩士 === 中原大學 === 生物科技研究所 === 106 === Zebrafish behavior can be defined as the coordination of the internal responses to a stimulus. There are several simple paradigms that have been developed to assess the zebrafish behavior including three-dimensional (3D) swimming behavior test, novel tank diving test, mirror biting test, social interaction test, shoaling test and predator escape assay. Generally, the measurement of behaviors in zebrafish requires different equipment and settings, and assessments are usually complicated and costly. In addition, the measurement of 3D swimming behavior in zebrafish relies on commercial software or requires sophisticated scripts, and depends on more than two cameras to capture the video. Moreover, zebrafish behavioral data is highly variable, even when collected within the same conditions. Thus, a high-throughput screening setup for zebrafish behavior tests by using open source software is needed in order to obtain high sample numbers. Here, we establish a simple and economic apparatus to detect 3D locomotion in adult zebrafish. Adult zebrafish were used since its capability to exhibit complex behaviors such as having strong social interaction with others, effective learning abilities, and portraying distinctive reactions to different environmental stimuli. This setup involves a single camera capture system that records zebrafish movement in a specially designed water tank with a mirror tilted at 45 degrees. The recorded videos are analyzed using idTracker, an open source program that is used for collecting motion trajectories by matching the feature on all crossing frames of objects based on appearance analysis, while spatial positions are calibrated by ImageJ software and 3D trajectories are plotted by Origin 9.1 software. This easy setting allowed scientists to track 3D swimming behavior of multiple zebrafish with low cost and precise spatial position, showing great potential for fish behavioral research in the future. In this report, we also presented a versatile setup that applied acrylic tanks into 5 vertical layers and 2 horizontal columns to perform multiple behavior assays such as novel tank diving test, mirror biting test, social interaction, shoaling and predator escape assay. In total, 10 different fish behaviors can be recorded simultaneously in a single video file, and the XY coordination of locomotion can be calculated by open source software of idTracker and ImageJ. This simple setting will allow scientists to assess behavioral endpoints of multiple zebrafish individuals, even other animals with high reliability and reproducibility.
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