Re: DsRed vs 2P

Posted by Craig Brideau on
URL: http://confocal-microscopy-list.275.s1.nabble.com/DsRed-vs-2P-tp5830859p5830871.html

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What wavelength did you have your Ti:Saph tuned to?  The 2p wavelength which
a dye prefers is often different from what you would expect.  Try tuning the
laser around its full range and see if you get better signal anywhere else.
 Do you have any filters in your microscope that could be blocking the laser
or causing other problems?

Craig


2010/12/13 Staffan Nyström <[hidden email]>

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> Dear list members,
>
> Has anyone any experience with visualizing DsRed using Ti:Saph (2 photon)?
> I have problems getting any decent fluorescence signal through between 700
> and 1000nm excitation and there is significant bleedthrough
> with EGFP. In the end I gave up since the "regular" 1 photon excitation way
> gave a much better S/N ratio with minimal bleed through.
>
> Any tips? We have an upright Zeiss LSM system and mostly use 40X 1.0 NA
> water dipping objectives.
>
> Best
>
> Staffan Nystrom
>
> PhD student
>
> MBB / Oncology & Pathology
>
> Karolinska Institutet
> Stockholm, Sweden
>