A Photoswitchable Solvatochromic Dye for Probing Membrane Ordering by RESOLFT Super-resolution Microscopy** |
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| Authors: | Dr Andrew T Frawley Kathryn G Leslie Dr Virginia Wycisk Dr Silvia Galiani Dr Dilip Shrestha Prof Christian Eggeling Prof Harry L Anderson |
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| Institution: | 1. Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford, OX1 3TA UK;2. MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS UK |
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| 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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| Keywords: | RESOLFT super-resolution microscopy solvatochromism membrane polarity lipid ordering |
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