7/23/2023 0 Comments Avidemux manualSince the major level of analysis is the individual behavioral tank and each raw video records four behavioral tanks simultaneously, we need to split the raw video into four separate quadrants. The raw recorded video is stored on a microSD card until transferred to a desktop computer for further processing. This gives us a lateral (side-wise) view of four tanks at one time. The camera is mounted to a tripod, positioned roughly 30 cm away from and at the same vertical height as the behavioral tanks. We record videos using a battery-powered GoPro in medium aperture mode at 30 frames per second. We hope to upgrade to infrared light sources in the near future as this will allow us to record behavior in complete darkness. We use enhanced lighting strategies, including a soft CFL backlight to increase the contrast between the fish and the white background in our videos. Recording high-quality video without disrupting the zebrafish’s natural behavior is a fine balance to strike between lighting conditions and leaving the fish undisturbed. Several additional software packages are necessary to analyze, summarize, and compare the coordinates to derive behavioral insights. Several individual software packages are necessary to edit and analyze the video files. These instructions are our current methods to understand zebrafish behavior by generating the coordinates of the swimming trajectory of each fish. While manual scoring by human visual assessment is sometimes necessary (e.g., for aggression or seizure assays), our main goal is to automate this analysis process as much as possible to decrease human-induced bias and increase the amount of useful data that can be derived from each experiment. Each recording of the behavioral experiments must be quantified in an objective manner to compare and understand subtle changes in the swimming behavior of the zebrafish following drug administrations or environmental manipulations. One major form of data for our lab is the recorded video files of our zebrafish behavioral experiments.
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