[Zbrafish] td-tomato vs mcherry as a red fluo reporter

W. Todd Penberthy via zbrafish%40net.bio.net (by wtpenber from mac.com)
Sat Dec 6 16:32:09 EST 2008


I have seen incredibly bright results with tandem dimer-tomato, but  
have not made a germ-line animal yet.

Animals harboring apparently single tdT (+) neurons were easily  
detected even with a 2.5 objective.

We did not find the signal to be too interfering to visualization of  
neighboring cells when expressed in just a few cells, however, when  
it was expressed in skin we could not see through tdT very well when  
using the TR filter.

Best,
W. Todd Penberthy

On Oct 21, 2008, at 11:22 AM, Philippe Herbomel wrote:

> Dear All,
>
> We are wondering about the best red fluorescent reporter to make  
> transgenic zebrafish with.
> If one does not want to target the protein to a particular  
> compartment (membrane or nucleus), DsRed2, the most used  
> presently,  is probably the best (very bright, with reasonable  
> maturation time).
> If otherwise, the most used presently is mCherry, which is quite (4- 
> fold) less bright.
> Now from the Tsien lab papers' data, td-Tomato appears both much  
> (over 5-fold) brighter than mCherry, and still compatible with  
> membrane or nuclear targeting;
> and indeed at least two papers then provided evidence that td- 
> tomato was not only far superior to mcherry but even to GFP (in  
> terms of brightness) as a reporter in transgenic mice.
> Now surprisingly, when a colleague of us injected either mcherry or  
> tdtomato mRNA into 1-cell zebrafish embryos,
> she found td-Tomato to be less bright than mcherry.
> So my question is: has anyone found this too, and/or tried to make  
> transgenic zebrafish with td-Tomato as reporter ? And what about  
> its actual brightness (and decay upon illumination) in zebrafish  
> embryos/larvae, as compared to the mcherry and dsred2 ?
>
> Thanks in advance for your testimonies on this.
> I will post a summary of your responses !
>
> Philippe Herbomel
>
>
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> Philippe Herbomel
> Unite Macrophages et Developpement de l'Immunite
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