Re: DsRed vs 2P

Posted by Haberman, Ann on
URL: http://confocal-microscopy-list.275.s1.nabble.com/DsRed-vs-2P-tp5830859p5831150.html

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Hi Staffan,

Which variant of DsRed are you using? Some of them are easy to excite between 850 - 950 nm. The immature state of DsRed has a pronounced green emission, but a variant created by Ben Glick is rapidly maturing and has very little green emission.

It can be purchased through Clontech. http://www.clontech.com/products/detail.asp?product_id=157258

Ann Haberman

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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

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Ann Haberman, PhD
Assistant Professor
Department of Laboratory Medicine
Yale University School of Medicine

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