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(Source: D. Frost and D. Sepich from Zebrafish Book 5th Edition)

1.   Rinse embryos in sterile, calcium-free Ringer's solution.
2.   Transfer about 10-100 embryos in a single drop of calcium-free Ringer's solution to the center of a tissue culture plate.
3.   Drop a sterile coverslip onto the embryos and smash them.
4.   Add more calcium-free Ringer's solution, remove the coverslip and suspend the cells by swirling the plate.
5.   Transfer the suspension to a sterile centrifuge tube and spin at 300 x g for 7 minutes.
6.   Remove most of the supernatant, triturate the pellet through a sterile narrow-bore Pasteur pipette to resuspend the cells and then add several ml of growth medium.
7.   Centrifuge, 300 x g, 7 minutes.
8.   Remove supernatant and resuspend in enough growth medium to make a final concentration of 15 fish per ml.

9.   Plate the cells and incubate at 28.5°C.