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Selective Electrochemical Bleaching of the Outer Leaflet of Fluorescently Labeled Giant Liposomes
Author(s) -
Perez Jimenez Ana Isabel,
Challier Lylian,
AïtYahiatène Eric,
Delacotte Jérôme,
Labbé Eric,
Buriez Olivier
Publication year - 2017
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201605786
Subject(s) - fluorescence , photobleaching , vesicle , biophysics , chemistry , fluorescence recovery after photobleaching , liposome , fluorescence microscope , confocal microscopy , electrochemistry , diffusion , analytical chemistry (journal) , membrane , chromatography , electrode , biochemistry , biology , optics , physics , microbiology and biotechnology , thermodynamics
Electrochemistry and confocal fluorescence microscopy were successfully combined to selectively bleach and monitor the fluorescence of NBD (7‐nitrobenz‐2‐oxa‐1,3‐diazole)‐labeled phospholipids of giant liposomes. Three types of giant unilamellar vesicles have been investigated, the fluorescent phospholipids being localized either mainly on their outer‐, inner‐, or both inner/outer leaflets. We established that only the fluorescent lipids incorporated in the outer leaflet of the vesicles underwent electrochemical bleaching upon reduction. The relative fluorescence intensity decay was quantified all along the electrochemical extinction through an original fluorescence loss in electrobleaching (FLIE) assay. As expected, the reorganization of the fluorescent phospholipids followed diffusion‐driven dynamics. This was also evidenced by comparison with fluorescence loss in photobleaching (FLIP) and the corresponding numerical model. The value of the lateral diffusion coefficient of phospholipids was found to be similar to that obtained by other methods reported in the literature. This versatile and selective bleaching procedure appears reliable to explore important biological and pharmacological issues.