[Zbrafish] Re: microinjections

Caroline Parkin via zbrafish%40net.bio.net (by caroline.parkin from sheffield.ac.uk)
Thu Mar 18 11:02:13 EST 2010


Dear Arash,

If I'm trying to make a transgenic line I inject into the cell or  
right underneath it, as we have a feeling that the sooner the DNA  
enters the cell the better the chance of incorporation into the  
genome. For MO or RNA we inject pretty much anywhere. However, as you  
can see from this video I made, the cytoplasmic streaming does seem to  
exclude the edges of the yolk somewhat. Maybe it's worth aiming for  
the centre of the yolk. Also the video shows that streaming stops at  
about the 16 cell stage, so it's not really worth injecting after that  
point.

Video: http://www.youtube.com/watch?v=nLuUPGIKQ4E

Best wishes,
Caroline Parkin
CDBG, University of Sheffield


On 11 Mar 2010, at 17:03, zbrafish-request from oat.bio.indiana.edu wrote:

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>   1. microinjectin into zebrafish embryo (Arash N.)
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> Message: 1
> Date: Wed, 10 Mar 2010 09:08:43 -0800 (PST)
> From: "Arash N." <arash321 from yahoo.com>
> Subject: [Zbrafish] microinjectin into zebrafish embryo
> To: zbrafish from magpie.bio.indiana.edu
> Message-ID: <132458.4604.qm from web110711.mail.gq1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Hello everyone,
>
> I just had a question regarding the procedures that people follow  
> regarding microinjections into Zebrafish embryos.
>
> Where inside the yolk do you typically aim to inject, where would  
> you ideally want to inject? Does it depend on what you are injecting?
>
> I understand that as long as it can be transported by the  
> cytoplasmic streaming it should work, does that mean that injecting  
> into the centre of the yolk would be sufficient?
>
> Are there any applications, where one would have to inject closer to  
> the cells, and where injecting only into the centre of the yolk  
> would not work?
>
> Overall, what I am trying to understand is what applications require  
> injections closer to the cells, and what applications work just fine  
> if you merely inject the material into the centre of the yolk.
>
> Any kind of feedback would be appreciated!
>
> Thank you
>
>
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