Figure 4
From: Analysis of vertebrate vision in a 384-well imaging system
Developed software and algorithms for automated analysis of zebrafish behavior in four adjacent 96-well plates. An ImageJ dialog box gives the user various options for automated analyses of behavior. The user is asked to outline a row of wells and the software will divide this area in 12 equal regions for measurements of individual wells. The software splits the color channels and uses one of the color channels, without visual stimuli, for automated image analysis. Subsequent images are subtracted, highlighting larvae that moved. This movement is counted over time as a measure of activity. In addition, the software compares the larval centroid to the well’s centroid to determine if a larva is located ‘up’ or ‘down’ in a well. Larval location is used to examine if moving visual stimuli drive the larvae up or down a well. A single results file of a 3-hour recording contains more than 10 million data points (15 measured or calculated values × 384 wells × 1800 images).
