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    • CommentAuthorjuruble
    • CommentTimeJul 11th 2007 edited
     

    This protocol explains how to: breed the zebrafish to obtain embryos.

    Protocol revised: 6/26/2007

    Protocol written by: Natasha Meyer

    1. In the afternoon (anytime after feeding) of the day before you want to breed, pair 1 male and 2 females in a breeding chamber (these hook into the larger tanks).  After you have filled the breeder bins with water, REFILL THE SYSTEM SUMP WITH WATER FROM THE CARBOYS. Water levels in the sump should be just under the white filter tray or in line with the black plunger.


    male and female

    1. At 8:00 AM the next morning (lights come on at 8:00), go down to the zebrafish facility to mate fish.
    2. Fill the plastic holding tanks 3/5 of the way up with fish water (E3).
    3. Transfer breeding chambers into holding tanks. Move these fish to the "drawer room" (first room on the left when entering the animal care facility) and allow them to acclimate for a few minutes.
    4. Come back and remove the dividers.
    5. Fish should lay eggs in 30 minutes to 1 hour.
    6. Place fish back in their normal tanks and remove the breeding chamber from the holding tank while gently shaking the bottom to release any caught eggs.
    7. Check the holding tanks for any clear eggs. If there aren't any eggs, dump the water. If there are, fill a small dish with E3.
    8. Pour the water from the holding tank into a fish net to separate the eggs from the water. Place your hand on the reverse side of the net (not touching the eggs directly) and lightly dip your hand with the netting into the water to release the eggs.
    9. Place 40 eggs in a dish with E3 to separate.
    10. Check the next morning for dead eggs and remove (if left in they will infect and kill the fertilized eggs). Dead eggs look clear with a white cloud inside.

    zf development

    Pictures from:

    http://www.fac.org.ar/scvc/llave/pediat/diaz/diazf9.jpg

    http://www.cas.vanderbilt.edu/bioimages/animals/danrer/zfish-devel.htm

    • CommentAuthorkalayman
    • CommentTimeJan 31st 2010
     
    From Kelli Carroll '09: I learned a couple breeding tricks for the fish that might come in handy for you guys, including leaving them in the breeding bins overnight to concentrate the pheremones (instead of in the system where water is circulated) and tipping the inner part of the container up so that it is at a slope like the shoreline. Do this in the morning when they are ready to lay.  Apparently in the wild they lay eggs using the shoreline, so this helps them.