Zebrafish Rheotaxis Assay: A Method for Measuring Orientation Behavior in Response to Different Water Flow Rates
筆記録
– First, carefully transfer the fish に the swimming tunnel and switch on the water pump. Let the fish acclimate for one hour に reduce its stress level. Fish tend に display positive rheotaxis or orienting upstream で flowing water. Using ビデオ analysis and a modeling software program, track the time and fish movement by marking its eye position at each upstream に downstream turn with a dot, and at each downstream に 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 に the total, thereby, quantifying the fish's orientation behavior. The example protocol is part of an experiment に 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 に a gas Tesla meter inside the tunnel and verify the voltage necessary に obtain the chosen magnetic field value along the major axis of the tunnel. Then disconnect the probe and connect the flow tunnel に the swimming apparatus. To set up a magnetic field with three dimensional magnetic field manipulation, turn on the computer, digital に analog converter, and coil drivers and set the chosen magnetic field on the xy and z-axis, then place the tunnel で the center of the Helmholtz pairs set. And the magnetic field of choice has been set up, select one に five fish for the rheotactic test. Transfer the fish で the tunnel, set the flow rate で the tunnel に 4.5 liters per minute and turn on the pump.
– Be sure に 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 で 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 に the chosen experimental protocol.
