title: Alternating Laser Excitation for Solution-Based Single-Molecule FRET.
abstract: Single-molecule fluorescence resonance energy transfer (smFRET) has been widely applied to the study of fluorescently labeled biomolecules on surfaces and in solution. Sorting single molecules based on fluorescent dye stoichiometry provides one with further layers of information and also enables "filtering" of unwanted molecules from the analysis. We accomplish this sorting by using alternating laser excitation (ALEX) in combination with smFRET measurements; here we describe the implementation of these methodologies for the study of biomolecules in solution.
authors.forename: Achillefs
authors.initials: A
authors.lastname: Kapanidis
authors.forename: Devdoot
authors.initials: D
authors.lastname: Majumdar
authors.forename: Mike
authors.initials: M
authors.lastname: Heilemann
authors.forename: Eyal
authors.initials: E
authors.lastname: Nir
authors.forename: Shimon
authors.initials: S
authors.lastname: Weiss
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes; D007834:Lasers; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins