[Zbrafish] dechorionating embryos

ESanders via zbrafish%40net.bio.net (by eriksanders from wustl.edu)
Mon Sep 19 16:10:34 EST 2011

You can manually dechorionate using two pair of very fine forceps, or use a
chemical method that takes a little practice but is vastly superior when you
need to deal with lots of embryos. See below SOP.

Pronase Dechorionation: 

The bleach treatment makes the chorion harder and the embryos will not be
able to hatch on their own.  Therefore they must be manually dechorionated
with forceps, or with pronase .

Solutions Required:
Pronase 20mg/ml  (Sigma P-5147)

1)	add 1mL of Pronase to a Petri dish of up to 250 embryos (in egg water)
for exactly 3 minutes
2)	pour off Pronase containing egg water and refill with autoclaved egg
water. Repeat this step for a total of 3 washouts.
3)	Refill dish with autoclaved egg water.

Some embryos will have hatched immediately, others may require 24-hrs to
hatch, or can be hatched by gently swirling or pipetting up and down with a
transfer pipette.

Good Luck,

Erik Sanders

Noopur wrote:
> Hi,
> Can anyone tell me the easiest way of dechorionating early stage
> zebrafish embryos without disrupting the yolk?
> Thanks in advance,
> With Best wishes
> Noopur
> _______________________________________________
> Zbrafish mailing list
> Zbrafish from net.bio.net
> http://www.bio.net/biomail/listinfo/zbrafish

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