title: Aligning the μs-ALEX Setup.
abstract: To achieve single-molecule sensitivity and thus have the ability to detect single diffusing fluorophores, careful alignment of the microsecond-alternating laser excitation (μs-Alex) setup is crucial. The following protocol describes routine alignment for 2c-ALEX (532 nm/635 nm) with spectral windows G(550-620)R(650-750).
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