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Fixed 4 samples for confocal microscopy.

  • 10 days, jelly dissolved with 5 min (stirring) + 5 min (while dissecting) 5% cysteine.
  • 10 days without cysteine.
  • 21 days, jelly dissolved with 10 min 10% cysteine.
  • 21 days without cysteine.

10% cysteine damaged the juveniles and some died. This concentration is too high, even with short times. 5% cysteine worked well with the earlier stages and it was much easier to dissect the eggs off the bundles afterwards (compared to the non-treated samples). The only thing is to see if the cysteine at this concentration damaged the embryos (or could affect the in situs).

Relaxation with MgCl2 is not working so well, the trunk of the juveniles from the 21 days samples were completely retracted. Or maybe try for a longer time (Maslakova2009 waited 10-15 min until they were completely relaxed). Also, they use poly-l-lisine treated slides.