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Cammer, Michael-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/advice-with-far-red-probe-to-distinguish-from-mRFP-mCherry-tp7591381.html
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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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