Zebrafish Avoidance and Thigmotaxis Assay: A High-Throughput Method to Examine Larval Behavior in Response to Aversive Visual Stimulus

Abstract

Source: Richendrfer, H., et al. Automated High-throughput Behavioral Analyses in Zebrafish Larvae.  J. Vis. Exp. (2013)

The video describes the avoidance and thigmotaxis assay of zebrafish larvae. The video examines the behavior of zebrafish larvae in response to visual aversive stimulus.

Protocol

 1. Image Capture Up to 20 larvae can be placed in each lane of the plates. Typically 5 larvae per lane are used to facilitate the most accurate tracking of swim speed and to reduce the number of larvae that are needed per experiment. The lanes can be filled with egg water with or without pharmaceuticals or toxicants depending upon the experiment. However, lanes should not be filled up all the way until they are placed in the imaging cabinets; this will prevent overflow. For consistency, the larvae shoul…

Representative Results

Figure 1. Imaging Cabinets. Imaging cabinets were specially built in our laboratory and used for high-throughput behavioral analyses. A 15 megapixel digital camera was attached to the top of the cabinet facing downwards in order to gather time lapse images of the larvae in multilane plates placed on top of the screen of a laptop. Between the plates and the screen there is a plastic diffuser that is used to preve…

Materials

Imaging Cabinets	WoodCraft Towers
Agarose	Fisher	BP1356-100
Digital camera	Canon EOS	Rebel T1i
Laptops	Acer Aspire		Any is good as long as it has a 15.6 in. LCD screen with 1366 x 768 pixel resolution and a brightness of 220 cd/m2.
One well plates	Fisher	12-565-493
PowerPoint 2010	Microsoft
Plastic diffuser	Pendaflex	52345