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We have set up a practical with students where we unmix AF555 and AF568 on a Zeiss LSM780. It works well as long as the intensities of the two fluorophores are comparable. 

Best regards,
Laure



________________________________________
From: Confocal Microscopy List [[log in to unmask]] on behalf of Craig Brideau [[log in to unmask]]
Sent: 07 April 2014 21:07
To: [log in to unmask]
Subject: Re: Spectrally unmixing mCherry and tdTomato

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We've unmixed Calcium Green and Newport Green, (Nikon C1Si, 32 channels) so
if your algorithm is up to the task and you have sufficient spectral
resolution, you should be fine.

Craig


On Mon, Apr 7, 2014 at 12:55 PM, G. Esteban Fernandez <
[log in to unmask]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> 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 doable
> on my LSM 710, but I wanted to check...has anyone actually spectrally
> unmixed mCherry and tdTomato successfully?
>
> Thanks,
> Esteban
>