(big) trouble after injection
cousin at ensam.inra.fr
Mon Aug 12 09:52:46 EST 2002
I am having big trouble with injected zebrafish embryos : they all
die before reaching 24h.
In more details, at 24h, i only find small white balls in the plates,
sometimes they have the size of yolk, sometimes, they are
smaller. Anyway most of the time this is all remaining my
embryos. I really have no idea of what happened during the nigth
because the day after injection development seems to be normal,
gastrulation occurs and segmentation begins... In addition some
(very rare) times i obtain 24h embryos expressing transgene...
What i do :
eggs collected at light onset eggs dechorionated with pronase
during 2-4 minutes and rinse gently 3-5 times in 0.3x danieau
injected at 2-4 cells stage sorted in the afternoon to remove
unfertilized eggs and abnormally developing embryos (small %
most of the time)
That's it !
Does someone have an idea ?
Please this would be a great help, because i tried several strategies
(i tested :
embryo densities leaving embryos on the injection plate or
changing plate (in this case changing to other agarose coated- or
glass plate) injected or not injected...)
All the best,
INRA - DCC tel. + 33 (0)4 99 61 28 14
2, place Pierre Viala fax + 33 (0)4 67 54 56 94
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