http://confocal-microscopy-list.275.s1.nabble.com/advice-with-far-red-probe-to-distinguish-from-mRFP-mCherry-tp7591381p7591385.html
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> Hi Michael,
>
> We use miRFP670 fairly regularly with 633 nm excitation and have not
> observed significant overlap with mCherry. We typically increase the laser
> power for miRFP670 compared to that for CFP, YFP or mCherry. Incubation
> with biliverdin considerably increases the fluorescence of iRFP670 as
> nicely documented in this paper below, and I can confirm that the increase
> in fluorescence with biliverdin is quite significant.
>
>
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5519290/>
> I hope this helps.
>
> Thanks
> Abhi
>
> Abhijit Deb Roy, PhD
> Cell Biology Department
> Johns Hopkins Medical Institute
> Baltimore, MD
>
>
>
> On Fri, Oct 9, 2020 at 2:37 PM Cammer, Michael <
>
[hidden email]> wrote:
>
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> >
> >
> >
> > A lab is asking us for advice with a far red fluorescent probe. They are
> > already using Dapi, GFP, and mCherry or mRFP and want to add a fourth
> > probe. They suggested E2-Crimson.
> >
> > I looked up E2-Crimson on FP Base (a great website) and found that we
> > could probably distinguish it well from mCherry by sequential excitations
> > and clipping some of the emission spectra if controls showed bleed
> > through. However, with a 633 nm laser excitation efficiency would only
> be
> > 40%.
> >
> > I was thinking something more redshifted would be preferable and found
> > miRFP670 which is reported to have 91% excitation efficiency with our
> > laser, but only half as bright, so it might be a wash.
> >
> > Any advice very much appreciated regarding:
> > Does anyone have experience with ER2-Crimson &/or miRFP670?
> > Is it as simple to separate from mCherry or mRFP as I expect based on
> the
> > reported ex and em data?
> > Any other suggestions for far red FPs?
> >
> > Thank you!!
> >
> > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> > NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY
> > 10016
> > Office: 646-501-0567 Cell: 914-309-3270
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