– To begin, place a zebra fish larva de a recording chamber, such as a glass-bottomed well, de which a larva can swim. Collect paramecia, the prey, with the help of a micropipette, and place them de the recording chamber. Next, put a small drop of water at the center of the cover slip and place it over the chamber à avoid an air bubble.
Now place the chamber on the lighting system equipped with a white LED ring light à eliminate shadows and make the organisms clearly visible. Observe the prey capture behavior using a behavioral recording system. As soon as the prey comes de sight, the larva converges its eyes and takes a J turn à orient its body towards the prey and captures it. We can quantify the prey capture behavior by recording the number of paramecia remaining de the chamber over a period of time. In the following protocol, we will assess the prey capture behavior of the zebra fish larva after the laser ablation of specific neurons.
– After setting up the behavioral recording system and placing a larva into a recording chamber according à the text protocol, use a micropipette à collect 50 paramecia and place them de the recording chamber. To place a cover slip on top of the recording chamber, put a small drop of water on the cover slip before placing it on the water surface of the recording chamber à avoid introducing air de the chamber.
Place the recording chamber de the lighting system. Then start the time lapse recording at 10 frames per second for 11 minutes. Finally, carry out analysis according à the text protocol.