A photoswitchable solvatochromic dye for probing membrane ordering by RESOLFT super-resolution microscopy

Authors:

Andrew Frawley, Kathryn Leslie, Virginia Wycisk, Silvia Galiani, Dilip Shrestha, Christian Eggeling, Harry Anderson

Keywords:

Vesicles, Membranes, RESOLFT

Abstract:

A switchable solvatochromic fluorescent dyad can be used to map ordering of lipids in vesicle membranes at a resolution better than the diffraction limit. Combining a Nile Red fluorophore with a photochromic spironaphthoxazine quencher allows the fluorescence to be controlled using visible light, via photoswitching and FRET quenching. Synthetic lipid vesicles of varying composition were imaged with an average 2.5-fold resolution enhancement, compared to the confocal images. Ratiometric detection was used to probe the membrane polarity, and domains of different lipid ordering were distinguished within the same membrane.

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