Reply to: dsred vectors intensity

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Fri Feb 11 11:35:17 EST 2000


Holger Fehr wrote:

> Dear Corinne,
>
> I my personal experience, using co-expression of similar or identical GFP-
> or dsRed-fusion proteins and analysis in a standard UV-microscope with
> filter sets for blue (485 nm) or green (530 nm) excitation, i found a very
> nice signal by the dsRFP using the filter for "red" dyes. The dsRed signal
> induced by "blue" light is detectable but significantly weaker, with a
> significant higher background (probably due to autofluorescence). The dsRed
> gave a higher resolution picture (e.g. for the "spotted" expression of
> cytokine receptors on the cell surface) with lower background compared to
> the corresponding GFP-constructs.
>

Hi Hoger,
I´ve also used the dsRed (RFP) and GFP in cotransfections, and found that RFP
is much weaker than GFP. We haven´t seen a higher background with RFP, but
signal to background was less favourable for photography then for GFP (using a
narrow-band filter for green fluorescence). With a standard filter set for
FITC, signal to background isn´t better for GFP than for RFP. I guess it will
be an improvement to use a narrow-band filter for red, optimized for RFP, but
I haven´t seen them on the market yet.

Frank





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