title: A photoprotection strategy for microsecond-resolution single-molecule fluorescence spectroscopy.
abstract: Time resolution of current single-molecule fluorescence techniques is limited to milliseconds because of dye blinking and bleaching. Here we introduce a photoprotection strategy that affords microsecond resolution by combining efficient triplet quenching by oxygen and Trolox with minimized bleaching via the oxygen radical scavenger cysteamine. Using this approach we resolved the single-molecule microsecond conformational fluctuations of two proteins: the two-state folder α-spectrin SH3 domain and the ultrafast downhill folder BBL.
authors.affiliation: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
authors.forename: Luis A
authors.initials: LA
authors.lastname: Campos
authors.forename: Jianwei
authors.initials: J
authors.lastname: Liu
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.forename: Ravishankar
authors.initials: R
authors.lastname: Ramanathan
authors.forename: Douglas S
authors.initials: DS
authors.lastname: English
authors.forename: Victor
authors.initials: V
authors.lastname: Muñoz
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008958:Models, Molecular; D038761:Photobleaching / Q000528:radiation effects; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D013049:Spectrin / Q000032:analysis / Q000737:chemistry; D013997:Time Factors
chemical_list: D013049:Spectrin