Reversibly photoactivatable fluorophores that are switched to an “on” state by visible light, and subsequently switch-off thermally, provide a level of control beneficial for imaging cellular features beyond the diffraction limit, when utilized as probes in super-resolution techniques such as Photoactivated Localization Microscopy (PALM) or REversible Saturable Optical Linear Fluorescence Transitions (RESOLFT). In this work, we introduce new derivatives of the bright, azetidine-containing Janelia Fluor dyes that can be activated by 405 nm light and switch back thermally. After activation, these dyes can then be excited using green or red light, emitting at ca. 550 nm and 650 nm. By incorporating either a HaloTag anchoring unit or membrane selective substituents, these photoactivatable fluorophores are capable of simultaneously and selectively labeling cellular sites of interest, allowing for multicolor super-resolution imaging.

Reversibly photoactivatable Janelia Fluor dyes, and their utility in imaging cellular features beyond the diffraction limit

253rd National Meeting (2017)

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