458 for GFP-RFP?

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mmodel mmodel
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458 for GFP-RFP?

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Dear Colleagues,

We need to buy optics to do mGFP-turbo RFP FRET, and it seems that excitation at 458 might give much less direct excitation of RFP with only little sacrifice in GFP excitation. But there might be some other consideration of which I am unaware. Has anyone had any good/bad experience using 458 line for GFP? Thanks in advance!

Mike Model
jerie jerie
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Re: 458 for GFP-RFP?

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Dear Mike,

because the 458 line  was a weak line of the Argon laser with less than 5%
of the total output power, for GFP excitation there were often limitations
to the excitation power, in particular when using low NA lenses and as a
consequence, also filter manufacturers offered less options including 458
in multiline dichroics.

So if you can provide a powerful enough laser -also to compensate for the
smaller excitation cross section- and suitable excitation optics, 458
should be well suited for GFPturbo excitation.

Greetings, Jens


On Tue, May 6, 2014 at 7:05 PM, MODEL, MICHAEL <[hidden email]> 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.
> *****
>
> Dear Colleagues,
>
> We need to buy optics to do mGFP-turbo RFP FRET, and it seems that
> excitation at 458 might give much less direct excitation of RFP with only
> little sacrifice in GFP excitation. But there might be some other
> consideration of which I am unaware. Has anyone had any good/bad experience
> using 458 line for GFP? Thanks in advance!
>
> Mike Model
>