4 replies to this topic

[kingsupercat]

if I'd like to trace a particular cell movement during zebrafish development,
could anyone give me good idea how can I achieve this purpose?
should I use specific fluorescent dye? won't that be diffusing if I inject into fish embryo?
i know somebody used the DMNB-caged fluorescence dextran before but Invitrogen is no longer providing it....

thanks for all of you who's giving me any kind of suggestion!!

[bob1]

inject them with something that will integrate into the DNA and be constituitively expressed (GFP?).

[kingsupercat]

Thanks for your reply, but I think the point is what if I'm monitoring the cell movement from "4 cell-stage" to 24 hpf? I guess the protein wouldn't be synthesized right after I inject the mRNA....

[shm267]

there are several techniques to achieve your purpose:
1. inject at late cleavage stage on random side of the embryos (with gfp, rfp, rldx, fldx,or whatever with fluorescence), and pick up the embryos at later developmental stages with fluorescence at the desired region.
2. using photoconversion strategy, such as caged dyes or Kaede-GFP, and label by UV irridation at the desired region.
3. transplant the labeled cells to the corresponding region of a recipient.

[basudec1509]

hello guys ...


its really nice and informative post....


i just liked it....


thanks for your information guys ...........





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