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how to trace single cell movement throughout gastrulation of zebrafish


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4 replies to this topic

#1 kingsupercat

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Posted 10 August 2009 - 08:07 AM

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!!
:(

#2 bob1

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Posted 10 August 2009 - 05:27 PM

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

#3 kingsupercat

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Posted 11 August 2009 - 02:52 AM

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....
:)

#4 shm267

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Posted 08 December 2009 - 08:05 PM

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.

#5 basudec1509

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Posted 01 February 2010 - 11:22 AM

hello guys ...


its really nice and informative post....


i just liked it....


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





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