Re: combination of 3 fluorescence protein reporters

Posted by Stephen Bunnell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/combination-of-3-fluorescence-protein-reporters-tp590861p590866.html

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Dear Xinyu,

    We have had good luck with that combination, as well as
mCerulean/mYFP/mCherry and mCerulean/mYFP/mStrawberry. Both of the latter
sets work well on our fixed dichroic Ultraview spinning disk system, which
has a custom built quad-pass dichroic that we acquired from Chroma. We have
no detectable bleedthough between these three channels using 442/514/568 nm
laser lines for excitation. By the book, Cherry looks like it should be
brighter and more stable, but Strawberry 'fits' our existing filter sets
(based on DsRed) much better.

    I had expected that mVenus should be much better that YFP, but in my
hands (and filter sets) it did not offer any significant advantages. It
actually bleached _faster_ than mYFP under constant 514nm illumination.

    Best regards,

    -Steve

****************************************************************************
Stephen C. Bunnell, Ph.D.
Assistant Professor
Tufts University Medical School
Department of Pathology
Jaharis Bldg., Room 512
150 Harrison Ave.
Boston, MA 02111

Phone: [phone]
Fax:   [phone]
Email: [hidden email]


On 11/8/07 10:06 AM, "Xinyu Zhao" <[hidden email]> wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear listers,
>
> One of our users wanted to image CFP and YFP labeled cells and would like to
> add a third fluorecence protein reporter later. I was wondering if anybody
> could make a recomendation on this third one.
>
> Anybody has ever used a  CFP/YFP/mCherry combination ?
>
> Thank you very much.
>
>
> Xinyu Zhao
> Biomedical Imaging Core Lab
> School of Medicine
> University of Pennsylvania
> Tel:  [phone]