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. Author manuscript; available in PMC: 2015 Feb 21.
Published in final edited form as: Chem Soc Rev. 2014 Feb 21;43(4):1044–1056. doi: 10.1039/c3cs60237k

Fig. 8.

Fig. 8

Self-healing fluorophores enable robust smFRET recordings at emission rates compatible with millisecond time scale imaging. Data were collected in Tris-polymix buffer using a wide-field TIRF illumination system as previously described142 at a frame rate of 66 Hz. (a) Cartoon illustrating the labeling strategy designed to probe inter-subunit rotation of the bacterial ribosome using smFRET. Low FRET reports on an unrotated ribosome conformation; high FRET reports on a rotated ribosome conformation. (b) A long-lived trace with emission rate of >100 photons per millisecond enabled by imaging with self-healing dyes. Molecular oxygen was depleted using the PCA/PCD system.