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

Treat embryos with bleach (0.5% solution for 2 min) before removing the chorions.  Decant the bleach solution and rinse with 10% sterile Hank's saline.  Remove the chorions with watchmaker's forceps or by soaking the embryos in Pronase (see Removing Embryos from Chorions, page 4.3).  Embryos that can move should be anesthetized by soaking in tricaine or by chilling on ice.  Embryos can be dissociated with 0 Ca{^}2+^up to about 16 hpf of development. Older animals should be dissociated with the protease treatment or by dissection. 

1 Comment

  1. May 31, 2012

    alby

    hi, I use this protocol to prepare embryos for immunofluorescent assays. The di…

    hi,

    I use this protocol to prepare embryos for immunofluorescent assays. The dissociation is Ok but I have a problem with the cellular attachment on a 96 microplate. In fact, cells in PBS are incubated 1 hour at room temp before treatment with 4% PFA. The problem is that cells don't like washes. Do you have an idea to facilate the attachment.

    thanks