Super-resolution imaging in live Caulobacter crescentus cells using photoswitchable EYFP

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Abstract
Previous work showed that the commonly used fluorescent protein EYFP can be bleached and reactivated. Exploiting this property allows super-resolution in vivo imaging of EYFP-labeled structures in living bacteria. Fölling et al., also in this issue, describe a related approach for super-resolution imaging using other ordinary fluorophores. The commonly used, monomeric EYFP enabled imaging of intracellular protein structures beyond the optical resolution limit ('super-resolution' imaging) in living cells. By combining photoinduced activation of single EYFP fusions and time-lapse imaging, we obtained sub–40 nm resolution images of the filamentous superstructure of the bacterial actin protein MreB in live Caulobacter crescentus cells. These studies demonstrated that EYFP is a useful emitter for in vivo super-resolution imaging.