Creating an Effective Set-Up for Assessing Larval Zebrafish Movement

Science Center, Bent Corridor

Understanding the functional impact of environmental factors on neurodevelopment is an important but challenging topic to study. One prospective way to investigate these questions in vivo is to utilize model organisms useful for studying neurodevelopment, such as zebrafish. Analysis of larval zebrafish swimming patterns following embryonic exposure to neurotoxins may provide key insights into any associated functional changes in neurodevelopment and mirror effects of these toxins in human neurodevelopmental disorders. Here, we sought to implement a protocol for the tracking and analysis of larval zebrafish using established MATLAB programs, LSRtrack and LSRanalyze. By loading fish into multi-well plates and recording swimming behavior over a predetermined period of time, we can track fish movement and quantify neurobehavioral phenotypes. Additionally, the program records errors in its ability to track fish, such as its inability to detect fish in their respective wells or the presence of well walls. We report here the results of optimization experiments used to determine the most accurate and replicable set-up for movement recording and analysis. Variables such as imaging orientation (camera above vs. inverted), well solution volume, and video software filters were examined. Unexpectedly, imaging with the camera above the plate was problematic due to issues with solution meniscus and well wall visibility. An inverted set-up was found to be the best arrangement, where the camera rests at the base of a platform and captures the bottom of the well plate lit from above by a light box.