Zebrafish Rheotaxis Assay: A Method for Measuring Orientation Behavior in Response to Different Water Flow Rates
Transcrição
– First, carefully transfer the fish para the swimming tunnel and switch on the water pump. Let the fish acclimate for one hour para reduce its stress level. Fish tend para display positive rheotaxis or orienting upstream de flowing water. Using video analysis and a modeling software program, track the time and fish movement by marking its eye position at each upstream para downstream turn with a dot, and at each downstream para upstream turn with dot of a different color. Increase the water flow rate and repeat the trial. We can calculate a rheotactic index by comparing the time spent facing upstream para the total, thereby, quantifying the fish's orientation behavior. The example protocol is part of an experiment para separate the influences of water flow and magnetic fields on the orientation behavior of fish.
– To set up a magnetic field with one dimensional magnetic field manipulation, keep the tunnel open and switch on power unit and magnetometer. Place a magnetic probe connected para a gas Tesla meter inside the tunnel and verify the voltage necessary para obtain the chosen magnetic field value along the major axis of the tunnel. Then disconnect the probe and connect the flow tunnel para the swimming apparatus. To set up a magnetic field with three dimensional magnetic field manipulation, turn on the computer, digital para analog converter, and coil drivers and set the chosen magnetic field on the xy and z-axis, then place the tunnel de the center of the Helmholtz pairs set. And the magnetic field of choice has been set up, select one para five fish for the rheotactic test. Transfer the fish de the tunnel, set the flow rate de the tunnel para 4.5 liters per minute and turn on the pump.
– Be sure para close the outlet valve before filling the tunnel with water, and transfer the fish and open it before setting the flow rate.
– After the animals have acclimated de the swimming tunnel for an hour, start recording the behavior of the fish within the tunnel and begin the stepwise increase of the flow rate according para the chosen experimental protocol.
