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Hi Daniela,

First paragraph of your discussion states, " ... mCherry (based on our 
novel observation of mCherry photoconversion to a red-shifted species)"

where is data on this?

Is this referring to your observation,

"reference spectra for each FP (Figure 1A). Unexpectedly, we observed 
that the mCherry spectrum was dependent on the excitation wavelength, 
and that 594 nm excitation resulted in ~12-nm-peak red-shift compared 
with 561-nm excitation."

which is more likely to be instrument than protein related.

Are you going to add a blue FP?
Are you adding fusion proteins, ex. cyan-yellow (re: CY11.5, CyPet-YPet) 
with moderate to high FRET, enabling more colors (if using sequential 
scanning ... may also better take advantage of MP/NDDs)?
How about using plasma membrane targeting to outline the cell surfaces? 
This would leave a cleaner signal for taking advantage of my Tattletales 
concept to multiplex fluorescent reporters and live cells 
(http://works.bepress.com/gmcnamara/26/ - best so start with the Feb 5 
pdf and ppt).

Sincerely,

George


On 2/15/2013 1:22 PM, Daniela Malide wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> We have a recent publication in Blood -imaging 5 fluorescent proteins on a
> commercial Leica SP5 confocal system:
>
> Dynamic clonal analysis of murine hematopoietic stem and progenitor cells
> marked by 5 fluorescent proteins using confocal and multiphoton microscopy
>
> Daniela Malide, Jean-Yves Métais and Cynthia E. Dunbar
> Blood December 20, 2012 vol. 120 no. 26 e105-e116
>
> http://bloodjournal.hematologylibrary.org/content/120/26/e105.long
>
> I can email a pdf of the manuscript if interested -as require subscription to
> Blood.
>
> Daniela Malide
> Staff Scientist
> NHLBI Light microscopy core facility
>
>