title: Accurate fluorescence quantum yield determination by fluorescence correlation spectroscopy.
abstract: Here, we present a comparative method for the accurate determination of fluorescence quantum yields (QYs) by fluorescence correlation spectroscopy. By exploiting the high sensitivity of single-molecule spectroscopy, we obtain the QYs of samples in the microliter range and at (sub)nanomolar concentrations. Additionally, in combination with fluorescence lifetime measurements, our method allows the quantification of both static and collisional quenching constants. Thus, besides being simple and fast, our method opens up the possibility to photophysically characterize labeled biomolecules under application-relevant conditions and with low sample consumption, which is often important in single-molecule studies.
authors.affiliation: †AG Biophysik, I. Physikalisches Institut (IA), RWTH Aachen University, 52062 Aachen, Germany.
authors.forename: Daryan
authors.initials: D
authors.lastname: Kempe
authors.affiliation: ‡Molecular Biophysics, Institute of Complex Systems (ICS-5), Forschungszentrum Jülich, 52428 Jülich, Germany.
authors.forename: Antonie
authors.initials: A
authors.lastname: Schöne
authors.affiliation: †AG Biophysik, I. Physikalisches Institut (IA), RWTH Aachen University, 52062 Aachen, Germany.|‡Molecular Biophysics, Institute of Complex Systems (ICS-5), Forschungszentrum Jülich, 52428 Jülich, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Fitter
authors.affiliation: ‡Molecular Biophysics, Institute of Complex Systems (ICS-5), Forschungszentrum Jülich, 52428 Jülich, Germany.
authors.forename: Matteo
authors.initials: M
authors.lastname: Gabba
mesh_terms: D004058:Diffusion; D005453:Fluorescence; D016014:Linear Models; D018613:Microscopy, Confocal / Q000379:methods; D008956:Models, Chemical; D010735:Phosphoglycerate Kinase / Q000737:chemistry; D055668:Photochemical Processes; D011789:Quantum Theory; D012996:Solutions; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014364:Tryptophan / Q000737:chemistry
chemical_list: D012996:Solutions; D014364:Tryptophan; D010735:Phosphoglycerate Kinase