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We have a couple folks who have successfully used iFP1.4 in the far red 
(Cy5) channel on our scopes.  On our four line spinning disk confocal 
(405/491/561/640) we have successfully done 4-color imaging in mammalian 
cells using mTagBFP / EGFP / mCherry / iFP1.4.  Probably there are 
better combinations out there (iRFP is supposed to be better than 
iFP1.4) but this one works well.

Kurt

On 11/2/2011 3:10 PM, Cameron, Lisa wrote:
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>
> Hello -
>
> I was wondering what people's experience is with the far red FP variants - mPlum
> or mRasberry
> Roger Tsien's Nat. Methods paper from 2005 says mPlum is better. Have people
> successfully been using this FP?
> Would be great to hear recommendations/things to consider.
> And if one would also want to image eGFP and another FP in between, what would
> people recommend for the optimal combination of spectral separation but also
> fluor brightness/stability?
>
> I have a core user who is aiming to express all three - I realize this is a
> difficult task, just for cell health and transfection, but was hoping for FP
> choice recommendations.
>
> Thank you for any feedback -
> Feel free to contact me off list.
> Lisa
>
>
> ---------------------------------------
> Lisa Cameron, Ph.D.
> Director of Confocal and Light Microscopy Core
> Dana Farber Cancer Institute
> Boston, MA 02215
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