Novel Tank Diving Assay: A Method to Study Innate Stress Responses in Zebrafish

- To begin, place an infrared light behind the novel tank and a white acrylic sheet in between the tank and the light to diffuse the light and prevent glare. Now fill 3/4 of the tank with fish system water generated by reverse osmosis of tap water. Transfer one acclimatized fish from a holding tank to a beaker containing fish system water and let it rest for another 10 minutes. Next, gently pour the water and the fish into the novel tank.

Using a still frame video image to find the top, middle, and bottom areas of the tank by calculating the height of the tank and dividing it by 3, determine the time spent in each area and the total distance traveled using tracking software. An adult fish, when placed in an unfamiliar tank, usually shows a preference for the bottom half, due to stress. As the stress level reduces, it gradually swims to the top more often. Test at least 10 fish per experiment. In an example protocol, we will see the experimental setup for a novel tank diving test.

- Begin by filling a 250 milliliter beaker and at least two holding tanks with fish system water. On the morning of the test, transfer at least 10 adult zebrafish for each experimental condition into indiv