To begin, dissolve 10 grams of DMEM powder and 5.95 grams of HEPES in one liter of distilled water, and adjust the pH of the media to 6.91. Aliquot the solution into smaller containers and store them at four degrees Celsius. Then mix 88%DMEM medium with HEPES, 10%fetal bovine serum, and 2%penicillin streptomycin.
To prepare the zebra fish, set up a container or a tank with approximately two liters of water for four to five adult zebra fish. Using a fish net, transfer four to five adult zebra fish into the prepared tank. Prepare separate containers with water for anesthetizing and recovering the zebra fish.
After anesthetizing an adult zebra fish, use a plastic spoon to transfer the fish to a Petri dish lined with a wet paper towel, and place the dish under a dissecting stereomicroscope. Next, using fine forceps, gently tug on the anterior to the posterior direction to remove the scales. Place each individual scale into a separate 0.2 milliliter tube containing the scale culture medium working solution, and put the tubes inside an incubator set to 28 degrees Celsius.
After scale removal, carefully return the zebra fish to the recovery tank and monitor it. Once the zebra fish has resumed movement, return it to the regular rearing tank.
