Posted by
Tom Blanpied on
URL: http://confocal-microscopy-list.275.s1.nabble.com/what-fluorescent-proteins-are-working-best-for-precision-localization-microscopy-nanoscopy-in-your-a-tp7580157p7580172.html
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Hi,
We use mEos whenever possible. mEos2 seemed great to us, as we never saw
any effects of the small dimerization tendency. mEos3 does look to be at least
as good, and it does seem a bit brighter; it is our go-to probe now. We avoid
the bulk of the tdEos, but did think it was a good molecule.
PA-mCherry we thought was pretty terrific at first, but we no longer use it
unless we really need the separate GFP channel for something. It is
considerably dimmer in our hands than mEos (photons/frame and number of
frames till bleaching), and also seems to have a tendency to produce a smaller
population of photoactivated molecules. We've not tried to determine whether
this was due to expression level or folding or a non-activatable fraction, but it
does not seem to be due simply to the diminished ability to localize dim or
quickly bleached molecules.
PA-GFP and Dronpa we did a little bit with, but in general we have so much
autofluorescent green stuff in our relatively old cultured neurons that anything
in that spectral range will be unsatisfying. We finally managed to obtain some
PS-CFP2, but haven't yet measured much about it; we're hopeful it will be the
best of that range.
I'll be curious to hear comparisons of dendra2-type molecules.
Tom