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You can feed spectral info from both, excitation and/or emission into
unmixing algorithms.

Cheers,  Jens
http://br.linkedin.com/pub/jens-rietdorf/6/4a3/189/
Am 16.04.2014 11:41 schrieb "Wendy Salmon" <[log in to unmask]>:

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>
> We get good separation on our Zeiss 710 without unmixing simply by
> exciting the tdTomato at 514nm and mCherry at 561nm and imaging
> sequentially.  We have the spectral emission selection, so there's likely a
> slight shift in our detection wavelengths, too (I'll have to power up the
> system to check), but the biggest difference is the excitation efficiency.
>  This assumes similar fluorophore concentrations, of course.
>
> Good luck!
> Wendy
>
> ~~~~~~~~~~~~~~~~~~~~~~~
> Wendy Salmon
> Light Microscopy Specialist
> Whitehead Institute for Biomedical Research
> W.M. Keck Imaging Facility
> 9 Cambridge Center, Rm 447
> Cambridge, MA 02142
> c: 617-429-0158
> e: [log in to unmask]
> w: http://staffa.wi.mit.edu/microscopy/
>
>
> On Mon, Apr 7, 2014 at 12:55 PM, G. Esteban Fernandez <
> [log in to unmask]> wrote:
>
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> > http://lists.umn.edu/cgi-bin/wa?A0=3Dconfocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.=
>
> > *****
> >
> > Hi everyone,
> >
> > A user is planning an experiment where spectral separation of mCherry
> and=
>
> > tdTomato in the same mouse tissue will be needed (single photon).  I
> know=
>
> > that their peak emissions should be ~25 nm apart so this seems very
> doabl=
> e
> > on my LSM 710, but I wanted to check...has anyone actually spectrally
> > unmixed mCherry and tdTomato successfully?
> >
> > Thanks,
> > Esteban
> >
>