The article describes a behavioral method to assess stress in zebrafish. In response to stress, zebrafish tends to swim at bottom of a tank until stress is removed. The sample protocol describes stepwise method for performing the assay.
Protocol
1. Novel tank test setup Prepare a 250 mL beaker pre-filled with fish system water, and at least two holding tanks. On the morning of the test, transfer at least 10 test adult zebrafish to be used for each experimental condition (controls and experimental adults) from fish facility into a holding tank, transfer them to the behavior room, and allow them to acclimate for at least one hour. NOTE: A power analysis should be performed before experimentation, yet i…
Representative Results
Figure 1. Diagram of the novel tank setup. (A) Dimensions of the 1.8 L trapezoidal novel tank as seen from the recording side of the tank. (B) Diagram of the setup including positions of the infrared lights, camera, and barriers used to minimize human interference.
Materials
Camera
We use Point Grey Grasshopper3 USB camera with lens from Edmund Optics.
Infrared filter
Edmund Optics
Video Acquisition Program
Use programs such as Virtualdub or FlyCapture because the acquisition framerate can be set.
Infrared LED lights
Assay tank
Aquaneering
Part number ZT180
Size: M3 1.8 liter
Stand and clamp, or standard tripod for camera
250mL beaker
Tracking software
We use Ethovision XT 13 from Noldus Information Technology