title: A versatile dual spot laser scanning confocal microscopy system for advanced fluorescence correlation spectroscopy analysis in living cell.
abstract: A fluorescence correlation spectroscopy system based on two independent measurement volumes is presented. The optical setup and data acquisition hardware are detailed, as well as a complete protocol to control the location, size, and shape of the measurement volumes. A method that allows to monitor independently the excitation and collection efficiency distribution is proposed. Finally, a few examples of measurements that exploit the two spots in static and/or scanning schemes are reported.
authors.affiliation: Institut Fresnel, Aix-Marseille Université, Campus de St Jérôme, F-13013 Marseille, France. patrick.ferrand@fresnel.fr
authors.forename: Patrick
authors.initials: P
authors.lastname: Ferrand
authors.forename: Martina
authors.initials: M
authors.lastname: Pianta
authors.forename: Alla
authors.initials: A
authors.lastname: Kress
authors.forename: Alexandre
authors.initials: A
authors.lastname: Aillaud
authors.forename: Hervé
authors.initials: H
authors.lastname: Rigneault
authors.forename: Didier
authors.initials: D
authors.lastname: Marguet
mesh_terms: D000465:Algorithms; D000818:Animals; D019556:COS Cells; D002138:Calibration; D002522:Chlorocebus aethiops; D049452:Green Fluorescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D012235:Rhodamines / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D018800:Thy-1 Antigens / Q000378:metabolism; D013997:Time Factors
chemical_list: D012235:Rhodamines; D018800:Thy-1 Antigens; C026188:rhodamine 6G; D049452:Green Fluorescent Proteins