title: Reduced dyes enhance single-molecule localization density for live superresolution imaging.
abstract: Cell-permeable rhodamine dyes are reductively quenched by NaBH4 into a non-fluorescent leuco-rhodamine form. Quenching is reversible, and their fluorescence is recovered when the dyes are oxidized. In living cells, oxidation occurs spontaneously, and can result in up to ten-fold higher densities of single molecule localizations, and more photons per localization as compared with unmodified dyes. These two parameters directly impact the achievable resolution, and we see a significant improvement in the quality of live-cell point-localization super-resolution images taken with reduced dyes. These improvements carry over to increase the density of trajectories for single-molecule tracking experiments.
authors.affiliation: Laboratory of Experimental Biophysics, Institute of Physics of Biological Systems, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland).
authors.forename: Lina
authors.initials: L
authors.lastname: Carlini
authors.forename: Alexander
authors.initials: A
authors.lastname: Benke
authors.forename: Luc
authors.initials: L
authors.lastname: Reymond
authors.forename: Gražvydas
authors.initials: G
authors.lastname: Lukinavičius
authors.forename: Suliana
authors.initials: S
authors.lastname: Manley
mesh_terms: D001894:Borohydrides / Q000737:chemistry; D002470:Cell Survival; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D015394:Molecular Structure; D010084:Oxidation-Reduction; D012235:Rhodamines / Q000032:analysis / Q000737:chemistry*; D014407:Tumor Cells, Cultured
chemical_list: D001894:Borohydrides; D005456:Fluorescent Dyes; D012235:Rhodamines; C025364:sodium borohydride
keywords: fluorophores; live-cell imaging; rhodamine; single-molecule studies; superresolution microscopy