Zebrafish Core Facility

Protocols

Bleaching Protocol

Embryos to be raised should be bleached during gastrula stages (will still hatch on their own) or by day 1 (you will need to remove chorions by treating with pronase or manually) of development follows:

  1. Bleach Solution- Label Bottle with date prepared. Can be kept for up to one week.
    • 500mL Egg Water
    • 350 uL Bleach
  2. Soak embryos in Bleach Solution for exactly 5 minutes. Trying to blach more than two lines at once will make it difficult to be consistent and accurate
    • Pour off original egg water and add bleach solution until dish is at least half filled
    • Gently swirl dish so that the embryos are evenly seperated
  3. Rinse embryos in Egg Water for 5 minutes
    • Pour off bleach solution: remove all bleach by using a pipette
    • Add egg water until dish is at least half filled
    • Gently swirl dish so that embryos are evenly seperated
  4. Repeat steps #2 and #3 so that embryos are bleached and rinsed a total of TWO times
  5. Rinse embryos two more time in egg water for 5 minutes
  6. Divide embryos into dishes of no more that 40
    • Manually dechorionate or treat with pronase if you bleached on day 1 as they can no longer free themselves. (Add 10 uL of 30mg/mL pronase [thaw pronase from -20C and mix, as it does settle] to each petri dish and place into the incubator. Make sure embryos are not clumped together, which causes retardation, rather spread them throughout the dish. Keep pronase on ice, as it will digest itself as well as lose activity.

EMBRYOS WILL NOT HATCH WITHOUT PRONASE IF BLEACHED AFTER TAILBUD STAGE. CHECK TO MAKE SURE THAT YOUR EMBRYOS HATCH AND REMOVE THE PRONASE BEFORE PUTTING THE BABIES INTO THE FACILITY.

Contact Us

Albert Einstein College of Medicine

1300 Morris Park Ave
Chanin 523
Bronx N.Y. 10461  

E-mail: clinton.depaolo@einstein.yu.edu
Phone: 718.430.4210

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