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Separating the two proteins should not be too difficult if you have the right filter sets, or if you have a spectral microscope system. If you check the spectra (e.g. invitrogen spectra viewer which you can Google), you will see that GFP emission starts at ~ 500 nm, so a narrow CFP filter should give litttle or no GFP signal. Conversely, if you excite GFP at around 488 nm, there should be little or no CFP excitation.  Your institution probably has a Microscopy Core facility, so the best is to talk to them... I'm sure they can point you in the right direction.

--
Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024


http://www.fhcrc.org


On Aug 1, 2012, at 12:55 PM, Ramana Sidhaye wrote:

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> To join, leave or search the confocal microscopy listserv, go to:
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> 
> Hi,
>  One of my collaborators has a mouse endogenously expressing 2 proteins (both are expressed in the same cells), one coupled to CFP and the other to GFP. I am curious about the spectrum- how does one best evaluate both proteins in the same sample without concerns about cross-reactivity.
> 
> Thanks,
> Ramana SIdhaye