Zebrafish Husbandry

Egg Collection

Protocol for obtaining zebrafish eggs and growing them to use the larvae for behavioral assays. This protocol is normally done from 10:30am to 12:30pm. Users should not enter the fish room before 10:30am.

1. Wear appropriate PPE when in the fish room. Always wear gloves; booties (shoe covers), long gloves, and apron should be worn when needed.

2. Prepare collection trays by snapping the lids onto the glassware.

3. Place collection trays in each tank (1 for each normal-sized tank, 2 for larger tanks)

4. Arrange the “grass” so that it is floating up.

5. Leave the trays in the tanks for 2 hours.

6. Collect the trays. Make sure not to take any adult fish to the surface.

7. Use strainer to filter the contents of the collection tray.

   Tip

   

8. Move the eggs from the strainer to the collection tank(1).

   1. Collection tanks can be found in the back of the fish room, in a bottom rack neard the fridge.
   Tip

   Wash the strainer with a small amount of egg water to gather the eggs towards the middle of the strainer. This will make it easier to transfer them to the collection tank.

9. Place empty collection trays and lids in disinfecting solution for 10 minutes.

10. Rinse collection trays, lids, and strainer with hot water and place back in storage.

11. Place collection tank in incubator while clean-up is finished.

12. If the fish haven’t been fed yet (as outlined in the log sheet pasted outside the door), feed the tanks used for egg collection with Gemma as specified in each tank. Press the “feeding button” before feeding (located in the middle row of tanks, all the way to the right when facing the tanks).

    If you feed the fish, please place a note (e.g. sticky note, tape) on the tanks you've feed so they don't get fed twice.

13. Label the container (e.g. with tape) with your name, the date, and the strain of fish you've collected.

14. Log your collection in the log binder located next to the incubators. Make sure to record a use by and euthanize by date.

Increasing Yield

If you are struggling to get enough eggs for your experiments, follow the steps below:

• Place more containers.
• The day before you need to collect, feed the tanks with one extra cube of shrimp. You should do this after they have been fed their normal shrimp (as outlined in the log sheet pasted outside the door).

Collection Tank Clean-up:

A day after the collection, unfertilized eggs must be removed from the tank using a transfer pipette. Likewise, if there are too many eggs in one tank, the eggs must be split into another tank.