C. elegans Blastomere dissectie: een methode om de eierschaal te verwijderen en embryonale cellen te dissocieren

- For this dissection, use two micropipettes: one with an opening approximately the same width as an egg and one that is twice as wide, and a series of solutions organized in wells to facilitate rapid transfer between them.

First, place adult hermaphrodites into egg salt solution, which is osmotically balanced to prevent embryos from bursting. Cut the worms in half to release the developing embryos.

Then, identify two-cell stage embryos. These cells are the blastomeres formed by the division of the fertilized ovum. Use the micropipette with the larger opening to transfer an embryo to hypochlorite solution, bleach, which will begin to break down the protective eggshell.

After only 40 to 55 seconds, transfer the embryo to Shelton's growth medium, or SGM. Wash each embryo briefly in two wells of SGM to remove all traces of the bleach before placing them in a third well.

Now, use the smaller micropipette to repeatedly pipette the embryo providing the mechanical force to remove the eggshell and expose the blastomeres. Gently continue to pipette the embryo to separate the individual blastomere cells. In the following protocol, we will see this technique in action.

- Hold each end of a mi