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If possible, spectral unmixing may help.  As a random thought, even if you
don't have a spectral detector, you might be able to ratio a narrowband
filtered signal vs a broader filtered signal to figure out what is Pyrene
and what is NADH.  You would only need two detectors.

Craig


On Tue, Dec 13, 2011 at 12:34 AM, Cameron Nowell <
[log in to unmask]> wrote:

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> Hi  List,
>
> We are trying to image uptake of a Pyrene based dye by some cells using
> two-photon excitation (as we don't have a UV confocal system). The
> pyrene is excitable at 345nm and emits at 380nm. We did some trials with
> straight dye and found we can excite it quite well in the 740-780nm
> range. Turns out that is where mitochondria light up too due to the
> large amounts of NADH they have. Very nice staining, who needs
> mitotracker.
>
> Does anyone have any secrets on how to get around this?
>
> Cheers
>
> Cam
>
>
> Cameron J. Nowell
> Microscopy Manager
> Centre for Advanced Microscopy
> Ludwig Institute for Cancer Research Melbourne - Parkville Branch
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